01 Automated Protein Expression and Purification: ExiProgen Selection Guide 3 Protein Synthesis Kit ExiProgen EC Protein Synthesis Kit 4 ExiProgen EC-

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1 Automated Protein Expression and Purification: ExiProgen TM Manual Protein Expression and Purification: AccuRapid TM & MagListo TM Preparation of Template DNA for Protein Expression Protein Service Protein Expression and Purification Phone: Protein Synthesis Service Phone: Gene to Protein Service Phone:

2 01 Automated Protein Expression and Purification: ExiProgen Selection Guide 3 Protein Synthesis Kit ExiProgen EC Protein Synthesis Kit 4 ExiProgen EC-Maxi Protein Synthesis Kit 6 ExiProgen EC-Tagfree Protein Synthesis Kit 7 ExiProgen EC-Disulfide Protein Synthesis Kit 8 Protein Purification Kit ExiProgen His-tagged Protein Purification Kit 9 ExiProgen Dialysis Kit NEW 10 Protein Synthesis & Purification Instrument ExiProgen Go to M. Instrument & Devices

3 Selection Guide Overview 바이오니아에서는무세포단백질발현방법을 ExiProgen TM 에적용시킨다양한 kit 들을개발하여판매하고있습니다. 연구용효소등일반적인단백질을 ~ 100 μg 정도로합성할수있는 ExiProgen TM EC Protein Synthesis Kit, 대량으로 500 μg 내외의단백질을합성할수있는 ExiProgen TM EC-Maxi Protein Synthesis Kit, 이황화결합이 2 개이상인단백질을합성하는 ExiProgen TM EC- Disulfide Protein Synthesis Kit, His-tag 이제거된단백질을합성할수있는 ExiProgen TM EC-Tagfree Protein Synthesis Kit 을제공하고있습니다. 뿐만아니라, 재조합단백질이발현된 E. coli cell lysate 또는동물세포로부터목적단백질을정제할수있는 ExiProgen TM Histagged Protein Purification Kit, ExiProgen TM Dialysis Kit 도제공하고있습니다. 원리및제품에대한보다자세한내용은 ExiProgen TM Guide Book 을참조하시기바랍니다. ExiProgen TM Guide Book 은바이오니아홈페이지 ( 에서무료로다운로드받으실수있습니다. 3

4 ExiProgen EC Protein Synthesis Kit 빠른시간내에다종의단백질들의합성여부와활성을확인하고자할때 Experimental Data <Detection of color> <Detection of fluorescence> sfgfp RFP AcGFP sfgfp RFP AcGFP 바이오니아의 ExiProgen TM 장비를이용하여단백질을전자동으로합성하는 kit 로, Expression Cartridge 와 Purification Cartridge 로구성되어있습니다. 합성하고자하는 template DNA 를첨가한 kit 를 ExiProgen TM 에장착한후, Protocol no. 902 ( 약 6 시간 ) 를가동시키면전자동으로단백질이발현되고, his-tag 를이용하여정제가진행됩니다. 1 회반응으로최대 16 종의서로다른고순도의단백질을 100 μg 내외로얻을수있습니다. Fully Automatic: DNA-in-Protein-out Template DNA 1 μg/kb를사용하여약 6시간이내에고순도로정제된단백질을전자동으로얻을수있습니다. High Performance 세포독성으로인해세포배양으로합성이불가능한단백질의합성도가능합니다. High Purity and High Yield 각각의반응 well 당순도 90% 이상의단백질을 100 μg 내외로얻을수있습니다. Reproducibility Template DNA만첨가하면전자동으로발현, 정제가이루어져재현성이높습니다. Application 연구용및상업용효소를포함하는단백질의연구개발에사용될수있으며, 특히다수의 mutant screening 분석에적용할수있습니다. Figure 1. Detection of fluorescence emitted from fluorescent proteins synthesized with ExiProgen TM. This result indicates that these synthesized sfgfp, RFP, and AcGFP are functionally active. Left panel: Color of each protein elution samples detected with naked eyes. Right panel: Fluorescence from protein elution samples detected with UV illuminator. 66 kda 45 kda 29 kda 20.1 kda 14.4 kda 66 kda 45 kda 29 kda 20.1 kda 14.4 kda (well ) M E P E P E P E P E P E P E P E P (well) M E P E P E P E P E P E P E P E P Figure 2. Expression and purification of CAT. Reproducibility is seen in all 16 wells with no detectable variation between wells. Lane 1~16: Number of the well M: AccuLadder TM Protein Size Marker (Low) E: Expression sample P: Purification sample BIONEER 4

5 ExiProgen EC Protein Synthesis Kit M M2 66 kda 45 kda 29 kda 20.1 kda 14.4 kda 116 kda 97.4 kda 66 kda 45 kda 29 kda 20.1 kda 14.4 kda Figure 3. PAGE data of various proteins synthesized from various templates (Plasmid DNA-10~120 kda, PCR product-10~60 kda). Up to 16 types of proteins can be expressed and purified simultaneously with an average of over 90% purity. Amount of loading sample is 1/35 of total sample. M1: AccuLadder TM Protein Size Marker (Low) 1: CalmL3 (17.5 kda), 2: RNase H (20 kda), 3: DUSP 3 (22 kda) 4: CAT (24 kda), 5: AcGFP (29 kda), 6: EF-Ts (34 kda) 7: VF (45 kda), 8: Poly A polymerase (50 kda) 9: M-MLV RTase (75 kda), 10: BM3 (117 kda) M2: AccuLadder TM Protein Size Marker (Broad) 66 kda 45 kda 29 kda 20.1 kda 14.4 kda M E P Time(min) Base line BSA Negative control DUSP3 sample Figure 4. PAGE & enzyme activity data of DUSP3. Left panel: Expression and purification of DUSP3 (protein tyrosine phosphatase). M: AccuLadder TM Protein Size Marker (Low) E: Expression sample, P: Purification sample Right panel: DUSP3 synthesized with ExiProgen TM has an enzyme activity. DUSP3 phosphatase activity was measured by incubating with 500 μm 3-O-methylfluorescein phosphate (OMFP). (Reaction buffer: 100 mm Tris-HCl (ph 8.2), 40 mm NaCl, 1 mm DTT, 20% Glycerol) 166 kda 97.4 kda 66 kda 45 kda 29 kda 20.1 kda 14.4 kda M E P M NC1 NC2 Sample GAPDH (500 bp) Figure 5. PAGE & enzyme activity data of M-MLV RTase. Left panel: Expression and purification of M-MLV RTase. M: AccuLadder TM Protein Size Marker (Broad) E: Expression sample, P: Purification sample Light panel: Amplification of GAPDH of Human total RNA with M-MLV RTase was synthesized with ExiProgen TM. Lane M: 100 bp DNA Ladder Lane NC1: Negative control 1 (No-RNA) Lane NC2: Negative control 2 (No-enzyme) Lane Sample: 10 ng Human total RNA from HeLa cell Cat. no. Product Description K-7300 ExiProgen TM EC Protein Synthesis Kit, 16 rxns K-7301 ExiProgen TM EC Protein Synthesis Kit, 32 rxns K-7302 ExiProgen TM EC Protein Synthesis Kit, 96 rxns Related Product K-7400 ExiProgen TM ProXpress PCR Template Kit, 16 rxns K-2631 AccuPower ProFi Taq PCR PreMix, 96 tubes, 20 μl rxn/tube K-7350 pbivt Vectors Set-1 K-3035 AccuPrep Gel Purification Kit, 200 rxns K-3034 AccuPrep PCR Purification Kit, 200 rxns D bp DNA Ladder, 250 μl (135 ng/μl) D kb DNA Ladder, 500 μl (130 ng/μl) D-2020 AccuLadder TM Protein Size Marker (Low), 500 μl D-2010 AccuLadder TM Protein Size Marker (Broad), 500 μl A-5041 ExiProgen TM, Instrument 5

6 ExiProgen EC-Maxi Protein Synthesis Kit 500 μg 이상의단백질을손쉽게얻고자할때 Fully Automatic: DNA-in-Protein (in storage buffer)-out Template DNA 0.6 μg/kb를사용하여약 25 시간이내에고순도로정제된단백질용액을 storage buffer로치환된상태로회수할수있습니다. Parallel Processing 1~8종의단백질을동시에합성할수있습니다. High Purity and High Yield 각각의반응 well 당순도 90% 이상의단백질을최대 500 μg까지얻을수있습니다. Application 바이오니아의특허기술인 ExiProgen TM 장비의 SECF (Stepwise Exchange Cell-Free) 단백질합성법을이용하여전자동으로많은양의단백질을합성할수있는 kit 로, Expression Cartridge 와 Purification Cartridge 로구성되어있습니다. Template DNA 를첨가한 kit 를 ExiProgen TM 에장착한후, Protocol no. 903 ( 약 25 시간 ) 을가동시키면순도 90% 이상의단백질을반응 well 당최대 500 μg 까지합성할수있습니다. 또한단백질발현, 정제및투석까지모든반응이전자동으로이루어지며, 최종목적단백질은원하는 storage buffer 에용해된상태로회수할수있습니다. 연구용및상업용효소를포함하는단백질의연구개발에사용될수있습니다. Experimental Data Figure 1. Synthesis of various proteins. Proteins can be expressed and purified simultaneously with an average of over 90% purity. Amount of loading sample is 1/80 of total sample. M1: AccuLadder TM Protein Size Marker (Low) 1: CalmL3 (17.5 kda) 2: DUSP 3 (22 kda) 3: CAT (24 kda) 4: AcGFP (29 kda) 5: RFP(30 kda) 6: EF-Ts (34 kda) 7: VF (45 kda) 8: BM3 (117 kda) M2: AccuLadder TM Protein Size Marker (Broad) BIONEER Cat. no. Product Description K-7310 ExiProgen TM EC-Maxi Protein Synthesis Kit, 8 rxns Related Product K-7350 pbivt Vectors Set-1 K-3030 AccuPrep Plasmid Extraction Kit, 200 rxns K-3034 AccuPrep PCR Purification Kit, 200 rxns D bp DNA Ladder, 250 μl (135 ng/μl) D kb DNA Ladder, 500 μl (130 ng/μl) D-2020 AccuLadder TM Protein Size Marker (Low), 500 μl D-2010 AccuLadder TM Protein Size Marker (Broad), 500 μl A-5041 ExiProgen TM, Instrument 6

7 ExiProgen EC-Tagfree Protein Synthesis Kit Histidine Tag 이제거된원형의단백질을얻고자할때 Application 단백질의구조분석이나, 단백질의약품개발, 항원단백질합성에응용될수있습니다. Experimental Data Expression sample M Purification sample M kda 45 kda 29 kda 66 kda 45 kda 29 kda 바이오니아의 ExiProgen TM 장비를이용하여원형그대로의단백질을전자동으로합성할수있는 kit 로, Expression Cartridge 와 Purification Cartridge 로구성되어있습니다. Template DNA 를첨가한 kit 를 ExiProgen TM 에장착한후, Protocol no. 904 ( 약 25 시간 ) 를가동시키면, 목적단백질을 his-tag 이있는상태로발현시킨후 tag 을제거하여순도 95% 이상의목적단백질을 storage buffer 에용해된상태로회수할수있습니다. Protein Synthesis without His-tag Template DNA 0.6 μg/kb를사용하여약 25시간이내에 protease의처리에의해서 his-tag이제거된목적단백질을회수할수있습니다. Fully Automatic: DNA-in-Protein (in storage buffer)-out 목적단백질은 storage buffer에용해된상태로회수됩니다. Parallel Processing and High Purity 1~8종의단백질을동시에합성할수있으며, 순도 95% 이상의단백질을얻을수있습니다 kda 14.4 kda 20.1 kda 14.4 kda Figure 1. Expression and purification of various proteins. M: AccuLadder TM Protein Size Marker (Low) (D-2020, Bioneer) 1: DUSP3 (22 kda), 2: hgh (23 kda), 3: CAT (24 kda), 4: AcGFP (28 kda) Note) DUSP3: Dual specificity protein phosphatase 3, hgh: Human growth hormone 1. SDS-PAGE Gel Data 2. Western Blot Data (k Da) M Figure 2. Data of SDS-PAGE and Western blot with His-tag antibody. M: AccuLadder TM Protein Size Marker (Low) 1: Expression sample of pbivt-tev-acgfp 2: His tag-tev-acgfp (His-tagged sample) 3: AcGFP (His-tag free sample) Cat. no. Product Description K-7320 ExiProgen TM EC-Tagfree Protein Synthesis Kit, 8 rxns Related Product K-7350 pbivt Vectors Set-1 K-3030 AccuPrep Plasmid Extraction Kit, 200 rxns K-3034 AccuPrep PCR Purification Kit, 200 rxns D bp DNA Ladder, 250 μl (135 ng/μl) D kb DNA Ladder, 500 μl (130 ng/μl) D-2020 AccuLadder TM Protein Size Marker (Low), 500 μl D-2010 AccuLadder TM Protein Size Marker (Broad), 500 μl A-5041 ExiProgen TM, Instrument 7

8 ExiProgen EC-Disulfide Protein Synthesis Kit Disulfide Bond 가있는목적단백질을얻고자할때 Application Pharmaceutical protein, ScFv 합성및항체스크리닝에응용될수있습니다. Experimental Data 바이오니아의 ExiProgen TM 장비를이용하여이황화결합 (disulfide bond) 을가지고있는목적단백질을합성하는 kit 로, Expression Cartridge 와 Purification Cartridge 로구성되어있습니다. Template DNA 를첨가한 kit 를 ExiProgen TM 에장착한후, Protocol no. 905 ( 약 36 시간 ) 를가동시키면단백질발현, 정제및투석까지모든반응이전자동으로이루어지며, 최종목적단백질은 storage buffer 에용해된상태로회수할수있습니다. Protein Synthesis with Disulfide Bond Template DNA 1μg/kb를사용하여이황화결합을가지고있는단백질을합성할수있으며, 합성된단백질은고유의활성을나타냅니다. Fully Automatic: DNA-in-Protein (in storage buffer)-out 목적단백질은 storage buffer에용해된상태로회수됩니다. Parallel Processing and High Purity 1~8종의단백질을동시에합성할수있습니다. Figure 1. SDS- PAGE & enzyme activity data. Left Pannel: Expression and purification of ScFV and rtpa in ExiProgen TM M: AccuLadder TM Protein Size Marker (Low) 1: ScFV containing 2 disulfide bonds 2: rtpa containing 9 disulfide bonds * Amount of purification sample is 1/40 of total sample. Right panel: rtpa synthesized has an enzyme activity. 1: Synthesis sample using ExiProgen TM EC-Maxi Protein Synthesis Kit 2: Synthesis sample using ExiProgen TM EC-Disulfide Protein Synthesis Kit * rtpa activity was measured by incubating with chromogenic substrate S Cat. no. Product Description K-7330 ExiProgen TM EC-Disulfide Protein Synthesis Kit, 8 rxns Related Product K-7350 pbivt Vectors Set-1 K-3030 AccuPrep Plasmid Extraction Kit, 200 rxns K-3034 AccuPrep PCR Purification Kit, 200 rxns D bp DNA Ladder, 250 μl (135 ng/μl) D kb DNA Ladder, 500 μl (130 ng/μl) D-2020 AccuLadder TM Protein Size Marker (Low), 500 μl D-2010 AccuLadder TM Protein Size Marker (Broad), 500 μl A-5041 ExiProgen TM, Instrument BIONEER 8

9 ExiProgen His-tagged Protein Purification Kit His-tagged Protein 을정제하고자할때 Application His-tag 을갖는목적단백질이발현된 E.coli cell lysate 으로부터목적단백질을정제하는데사용될수있습니다. Experimental Data M CL P 45 kda 29 kda 바이오니아의 ExiProgen TM 을사용 (protocol no. 901, 약 2 시간 ) 하여재조합단백질이발현된 E. coli 세포파쇄액또는무세포단백질발현시료로부터 his-tag 을가지고있는목적단백질을고순도로정제하는 kit 입니다. Kit 내에는 Ni-NTA magnetic bead 와정제에필요한 buffer 가 pre-filled 되어있는 cartridge 가포함되어있습니다. Ni-NTA magnetic bead 는당사가개발한자성실리카입자로입자표면의니켈이온이단백질의 His-tag 과결합할수있어, 목적단백질을고순도로정제할수있습니다. Availability of Various Sample In vivo (E. coli 발현 ) 또는 in vitro ( 무세포발현 ) 로발현된재조합단백질의정제가가능합니다. Pre-filled Buffer Cartridge System 단백질정제에필요한모든버퍼가들어있는 pre-filled cartridge가제공되므로, 세포파쇄액또는무세포단백질발현시료만준비하면즉시사용이가능합니다. Parallel Processing and High Purity 1~16종의단백질을동시에정제할수있으며, 순도 90% 이상의단백질을얻을수있습니다 kda Figure 1. Purification of DUSP3 with ExiProgen TM. M: AccuLadder TM Protein Size Marker (Low) CL: Sample of recombinant E. coli cell lysate (DUSP3 expression) P: Sample of purification Related Product Cat. no. Product Description K-7220 ExiProgen TM His-Tagged Protein Purification Kit, 16 rxns K-7221 ExiProgen TM His-Tagged Protein Purification Kit, 32 rxns D-2020 AccuLadder TM Protein Size Marker (Low), 500 μl D-2010 AccuLadder TM Protein Size Marker (Broad), 500 μl A-5041 ExiProgen TM, Instrument 9

10 ExiProgen Dialysis Kit NEW 단백질의 buffer 를교환하고자할때 Application 단백질 buffer 교환, 단백질최적 buffer screening Experimental Data 바이오니아의 ExiProgen TM 장비를이용하여단백질을전자동으로 dialysis 할수있는 kit 입니다. 교환하고자하는 buffer를 kit내의 cartridge에넣어준후 ExiProgen TM 에장착하고, 이미설치되어있는 Protocol ( 약 13시간 ) 을가동시키면 buffer 교환이전자동으로이루어지며, 최종목적단백질을원하는 buffer에용해된상태로회수할수있습니다. Efficiency 적은양의 buffer를이용하여원하는 buffer로교환할수있습니다. Convenience 추가적인냉장시설없이저온에서 buffer를교환할수있습니다. Fully Automatic 시료로딩후전자동으로모든과정이진행됩니다. Parallel Processing 16종의단백질의 buffer를동시에교환할수있습니다. Figure 1. Glycerol 농도에따른효율비교 mm Tris-Cl (ph7.6), 100 mm NaCl, 0.2 mm EDTA, 0.05% NaN mm Tris-Cl (ph7.6), 100 mm NaCl, 0.2 mm EDTA, 0.05% NaN3, 10% Glycerol 3 50 mm Tris-Cl (ph7.6), 100 mm NaCl, 0.2 mm EDTA, 0.05% NaN3, 20% Glycerol 4 50 mm Tris-Cl (ph7.6), 100 mm NaCl, 0.2 mm EDTA, 0.05% NaN3, 30% Glycerol 5 50 mm Tris-Cl (ph7.6), 100 mm NaCl, 0.2 mm EDTA, 0.05% NaN3, 40% Glycerol 6 50 mm Tris-Cl (ph7.6), 100 mm NaCl, 0.2 mm EDTA, 0.05% NaN3, 50% Glycerol 7 PBS mm HEPES (ph7.6), 100 mm NaCl, 0.2 mm EDTA, 0.05% NaN3 Figure 2. Manual 와 automation 효율비교. Cat. no. Product Description K-7240 ExiProgen TM Dialysis Kit, 16 rxns Related Product D-2020 AccuLadder TM Protein Size Marker (Low), 500 μl D-2010 AccuLadder TM Protein Size Marker (Broad), 500 μl A-5041 ExiProgen TM, Instrument BIONEER 10

11 02 Manual Protein Expression and Purification: AccuRapid & MagListo Selection Guide 13 Protein Synthesis Kit AccuRapid Protein Synthesis Kit 13 AccuRapid Protein Expression Kit 14 AccuRapid Disulfide Protein Synthesis Kit NEW 16 Protein Purification Kit MagListo His-tagged Protein Purification Kit 17 MagListo Protein A, Protein G Kit 18

12 Selection Guide Overview 본제품군은 manual type 제품군으로, 용도에따라사용자가선택할수있도록단백질발현에서정제까지수행할수있는제품 (AccuRapid TM Protein Synthesis Kit, AccuRapid TM Disulfide Protein Synthesis Kit) 과, 단백질발현만수행하는제품 (AccuRapid TM Cell-Free/ Midi/Maxi Protein Expression Kit), 단백질정제만수행하는제품 (MagListo TM His-tagged Protein Purification Kit, MagListo TM Protein A, Protein G Kit) 으로나뉘어져있습니다. BIONEER 12

13 AccuRapid Protein Synthesis Kit The Easy Method for Protein Synthesis!! Application 연구용및상업용효소를포함하는단백질의연구개발에사용될수있습니다. Experimental Datata Expression sample Purification sample AccuRapid TM Protein Synthesis Kit 는 manual type 으로원하는단백질의발현을확인하고정제할수있는 kit 입니다. 본 kit 는 E. coli 무세포발현및 Ni-affinity 정제실험에사용되는모든구성품을포함하고있으며, 반응당정제된목적단백질을 100 μg 내외로회수할수있습니다. Figure 1. SDS-PAGE data of various proteins synthesized from various templates. M: AccuLadder TM Protein Size Marker (Broad) 1: CalmL3 (17.5 kda) 2: DUSP 3 (22 kda) 3: CAT (24 kda) 4: AcGFP (29 kda), 5: ERFP (31 kda) 6: EF-Ts (34 kda) 7: VF (45 kda) 8: BM3 (117 kda) High Simplicity and High Performance 세포를이용하지않고시험관튜브내에서손쉽게단백질을합성할수있으며, 기존의세포배양방법으로발현이불가능했던단백질도합성이가능합니다. Flexibility T7 발현시스템에서단백질발현이가능한다양한 template DNA 로부터원하는단백질의발현및정제여부를손쉽게확인할수있습니다. Compatibility ExiProgen TM EC Protein Synthesis Kit와호환이가능합니다. Cat. no. Product Description K-7280 AccuRapid TM Protein Synthesis Kit, 5 rxns Related Product K-3030 AccuPrep Plasmid Extraction Kit, 200 rxns K-3034 AccuPrep PCR Purification Kit, 200 rxns K-3035 AccuPrep Gel Purification Kit, 200 rxns D bp DNA Ladder, 250 μl (135 ng/μl) D kb DNA Ladder, 500 μl (130 ng/μl) D-2020 AccuLadder TM Protein Size Marker (Low), 500 μl D-2010 AccuLadder TM Protein Size Marker (Broad), 500 μl K-7350 pbivt Vectors Set-1 K-7400 ExiProgen TM ProXpress PCR Template Kit, 16 rxns 13

14 AccuRapid Protein Expression Kit Screening for Protein Expression Experimental Data 1. AccuRapid TM Protein Expression Kits 를이용한 AcGFP 단백질 (positive control) 발현 AccuRapid TM Protein Expression Kit 는 manual type 으로반응튜브내에서 template DNA 를이용하여 3 시간만에원하는단백질의발현을확인할수있는 kit 로서, E. coli extract 와 master mix 를포함하고있습니다. 이중 E. coli extract 에는 T7 RNA polymerase, ribosome 등이포함되어있고, master mix 는아미노산, rntps, 에너지원등이포함되어 template DNA 만있으면, 손쉽게목적단백질의발현을확인할수있습니다. Figure 1. Positive control protein expression data of AccuRapid TM Protein Expression Kits. (SDS-PAGE and Coomassie Brilliant Blue staining) M: AccuLadder TM Protein Size Marker (Low) (Bioneer Cat.no. D-2020) 1,3,5: Negative control(no-dna) 2,4,6: Positive control (pbivt-acgfp) Note) K μl rxn, K ml rxn, K ml rxn High Speed 3시간만에 DNA로부터단백질을발현할수있습니다. Flexibility T7 발현시스템에서단백질발현이가능한다양한 template DNA 로부터원하는단백질의발현여부를손쉽게확인할수있습니다. High Performance Host 세포에독성을나타내어기존의 cloning 방법으로발현이불가능했던단백질의발현이가능합니다. Compatibility 본 kit는 ExiProgen TM 단백질합성 kit류와호환이가능하며, template DNA의최적농도를찾는데사용할수있습니다. 2. 다양한단백질의발현 M kda 45 kda 29 kda 20.1 kda 14.4 kda PTP18 GFP CAT Application 연구용및상업용효소를포함하는단백질의연구개발에사용될수있습니다. Figure 2. Various protein expression data (SDS-PAGE and Coomassie Brilliant Blue staining) M: AccuLadder TM Protein Size Marker (Low) (Bioneer Cat. no. D-2020) 1,3,5: Negative control (No-DNA) 2: CAT (Chloramphenicol acetyl transferase) 4: GFP (Green fluorescence protein) 6: PTP1B(Protein tyrosine phosphatase 1B) BIONEER 14

15 AccuRapid Protein Expression Kit 3. Template DNA 의최적 DNA 농도 screening Figure 3. Determination of the optimal amount of template DNA for the expression of pbivt-gfp. (SDS-PAGE and Coomassie Brilliant Blue staining) M: AccuLadder TM Protein Size Marker (Low) (Bioneer Cat. no. D-2020) 1: Negative Control(No-DNA) 2: CAT (Chloramphenicol acetyl transferase) 3 ~8: pbivt-gfp(50, 100, 200, 400, 600, 800 ng) Cat. no. Product Description K-7250 AccuRapid TM Cell-Free Protein Expression Kit, 45 μl x 24 rxns K-7260 AccuRapid TM Midi Protein Expression Kit, 1 ml x 5 rxns K-7270 AccuRapid TM Maxi Protein Expression Kit, 10 ml x 1 rxn Related Product K-3030 AccuPrep Plasmid Extraction Kit, 200 rxns K-3034 AccuPrep PCR Purification Kit, 200 rxns K-3035 AccuPrep Gel Purification Kit, 200 rxns D bp DNA Ladder, 250 μl (135 ng/μl) D kb DNA Ladder, 500 μl (130 ng/μl) D-2020 AccuLadder TM Protein Size Marker (Low), 500 μl D-2010 AccuLadder TM Protein Size Marker (Broad), 500 μl K-7350 pbivt Vectors Set-1 K-7400 ExiProgen TM ProXpress PCR Template Kit, 16 rxns 15

16 AccuRapid Disulfide Protein Synthesis Kit NEW The Easy Method for Disulfide-bonded Protein Synthesis! Application Pharmaceutical protein, 다양한항체합성및항체스크리닝에응용될수있습니다. Experimental Data AccuRapid TM Disulfide Protein Synthesis Kit 는 manual type 으로이황화결합 (disulfide-bond) 을가지고있는목적단백질을합성하는 kit 입니다. 본 kit 는 E. coli 무세포발현및 Ni-affinity 정제실험에사용되는모든구성품을포함하고있으며, 이황화결합을가지고있는단백질을합성할수있습니다. Protein Synthesis with Disulfide-boned 이황화결합을가지고있는단백질을합성및정제할수있으며, 정제된단백질은고유의활성을나타냅니다. Flexibility T7 발현시스템에서단백질발현이가능한다양한 template DNA 로부터원하는단백질의발현및정제여부를손쉽게확인할수있습니다. Compatibility ExiProgen TM EC-Disulfide Protein Synthesis Kit와호환이가능합니다. Figure 1. Expression and purification of antibody fragment with AccuRapid TM Disulfide Protein Synthesis Kit. (12% SDS PAGE gel stained with Coomassie Blue R-250 and samples were loaded by 7.5% in one well). M: AccuLadder TM Protein Size Marker (Broad, Bioneer, D-2010) PC: positive control DNA (single-chain variable fragment) 1: VL (Variable domain of light chain) 2: VH (Variable domain of heavy chain) 3: LC (Light chain) 4: VH-CH1 (Variable & conserved domain 1 of heavy chain) 5: VL- Lκ(Variable domain of light domain & light kappa chain) 6: VH-Lκ(Variable domain of Heavy chain & light kappa chain) Cat. no. Product Description K-7290 AccuRapid TM Disulfide Protein Expression Kit, 5 rxns Related Product K-7330 ExiProgen TM EC-Disulfide Protein Synthesis Kit, 8rxn K-3030 AccuPrep Plasmid Extraction Kit, 200 rxns D bp DNA Ladder, 250 µl (135 ng/µl) D kb DNA Ladder, 500 µl (130 ng/µl) D-2020 AccuLadder TM Protein Size Marker (Low), 500 µl D-2010 AccuLadder TM Protein Size Marker (Broad), 500 µl K-7350 pbivt Vectors Set-1 BIONEER 16

17 MagListo His-tagged Protein Purification Kit Fast Micro Scale Purification of 6x His-tagged Protein Application 본 kit 는 manual type 으로 his-tag 을갖는목적단백질을정제하는데사용될수있습니다. Experimental Data MagListo TM His-tagged Protein Purification Kit 은 magnetic bead 로 his-tagged protein 을정제하는 kit 로서, Ni-NTA Magnetic Nanobead 와 his-tagged protein 정제에필요한 buffer 로구성되어있습니다. 6x histidine 을포함하는 target 단백질은 Ni- NTA Magnetic Nanobead 표면의 Ni 2+ 이온과특이적으로결합하여기타단백질로부터분리된후, 고농도의 free imidazole 처리시 bead 로부터용출됩니다. 평균직경 400 nm 의 Magnetic Nanobead 는표면적이넓고결합력이우수하여높은 binding capacity 를나타내며, 강력한자성력을가지고있어손실없이빠르게회수됨으로써실험시간을줄이고수율을높입니다. High Speed Ni-NTA Magnetic Nanobead를이용하여 his-tagged protein 을빠르고쉽게정제 High Performance SSMB (Spherical Shape Magnetic Beads) 에의한고순도 protein 획득 High Efficiency Crude cell lysate를이용한효율적인 screening Specification Mini scale Midi scale Maxi scale Preparation time ~ 5분 ~ 17분 ~ 26분 Elution volume 0.2 ml 2 ml 5 ml Binding capacity 30~40 mg his-tagged protein/1 g Ni-NTA beads Figure 1. Purification of 6x His-tagged protein. M: Protein broad marker (Bioneer, D-2010) 1: Loading sample 2: Unbound sample 3: Eluted sample (MagListo TM His-tagged Protein Purification Kit) Figure 2. Protein size effect on purification. M : Protein broad marker (Bioneer, D-2010) 1: Size 94 kda His-tagged protein 2: Size 67 kda His-tagged protein 3: Size 38 kda His-tagged protein 4: Size 27 kda His-tagged protein 5: Size 25 kda His-tagged protein Cat. no. Product Description K-7200 MagListo TM His-tagged Protein Purification Kit,1 ml x 5 Related Product TA AccuNanoBead TM Ni-NTA Magnetic Beads, size 400 nm, 0.5 g/25 ml D-2020 AccuLadder TM Protein Size Marker (Low), 500 μl D-2010 AccuLadder TM Protein Size Marker (Broad), 500 μl 17

18 MagListo Protein A, Protein G Kit Fast and Easy Purification of Antibody Application Antibody purification, Antigen purification, Immunoprecipitation, Protein-protein interaction, and Cell separation etc. Experimental Data (A) M (B) 66 kda 45 kda 29 kda 20 kda µg Human IgG/mg Beads Bioneer Competitor L Competitor Q MagListo TM Protein A Kit 와 MagListo TM Protein G Kit 는 magnetic bead 로항체를정제하는 kit 로서, Protein A 또는 Protein G Magnetic Nanobead 와항체정제에필요한 buffer 로구성되어있습니다. 각제품은고순도 (>95%) protein A 또는 G 가코팅된 magnetic silica bead 를사용하고있습니다. 평균직경 400 nm 의 Magnetic Nanobead 는표면적이넓고결합력이우수하여높은 binding capacity 를나타내며, 강력한자성력을가지고있어손실없이빠르게회수됨으로써실험시간을줄이고수율을높입니다. Fast Binding 강력한자성력은 bead의빠른회수율로실험을보다빠르고손실없이진행할수있습니다. Large Surface Area 평균직경 400 nm의 Magnetic Nanobead는표면적이넓고결합력이우수하여높은 binding capacity를보장합니다. Specificity 균일한구형의 nanobead로비특이적결합을감소시켰습니다. Figure 1. Comparison of protein binding efficiency of MagListo TM protein G Kit with competitor s kits. A) Upper and lower bands are heavy (HC) and light chain (LC) of purified human IgG in SDS-PAGE gel, respectively. M: Protein size marker (low), 1: Bioneer, 2: Competitor L, 3: Competitor Q B) Amount of bound IgG is quantified by UV-spectrophotometer. All reactions are carried out for 1 hr at room temperature. Human IgG is purchased from Sigma (Cat. no. I4506). 66 kda 45 kda 29 kda 20 kda M IgG M scfv Heavy chain Light chain 66 kda 45 kda 29 kda 20 kda Figure 2. Purification of IgG and scfv using MagListo TM Protein A Kit Specification Protein G Magnetic Nanobeads Protein A Magnetic Nanobeads Matrix Silica-coated Fe3O4 Silica-coated Fe3O4 Concentration 40 mg/ml (4% suspension) 40 mg/ml (4% suspension) Binding capacity > 30 μg Human IgG/mg beads > 20 μg Human IgG/mg beads BIONEER Cat. no. Product Description K-7710 MagListo TM Protein G Kit, 1 x 1 ml K-7720 MagListo TM Protein A Kit, 1 x 1 ml Related Product TA ~3 AccuNanoBead TM Protein G Magnetic Nanobeads, size 400 nm, 40 mg/1 ml, 200 mg/10 ml, 500 mg/25 ml TA ~3 AccuNanoBead TM Protein A Magnetic Nanobeads, size 400 nm, 40 mg/1 ml, 200 mg/10 ml, 500 mg/25 ml 18

19 03 Preparation of Template DNA for Protein Expression Overview 20 ExiProgen ProXpress PCR Template Kit 21 ExiProgen Circular PCR Template Kit NEW 23 pbivt Vector Set-1 25

20 Overview Overview ExiProgen TM 을이용하여목적단백질을발현하기위해서는무세포단백질발현용 template DNA 를먼저준비해야합니다. Template DNA 는 T7 promoter - Ribosome Binding Site (RBS) - Target gene (6x His-tag sequence 포함 ) - T7 terminator 의구조를가지고있어야하며, 그구조는아래의그림과같습니다. 또한 target gene 은 start codon 과 stop codon 을가지고있어야합니다. 그리고선택적으로 Ni-affinity 정제를하기위하여 5 말단또는 3 말단에 6x His-tag 을가지고있어야합니다. 발현벡터는바이오니아에서판매하는 in vitro translation 전용벡터인 pbivt 벡터 (Cat. no. K-7350) 에 cloning 하여사용할수있고, 그밖에도 pk7, pivex, pet 벡터등을사용할수있습니다. PCR product 로부터 cloning 을하지않고바로 template DNA 를제작하려면당사의 ExiProgen TM ProXpress PCR Template Kit (Cat. no. K-7400, 7401) 를이용하여제작할수있습니다. 많은종류의단백질을소량으로빠르게만들어서성능을평가해볼때매우유용한 kit 로서 genomic DNA, cdna 또는 T-vector 등에서간편하게 template DNA 제작부터단백질합성까지 1 일이내로진행할수있는장점이있습니다. 바이오니아에서는 template DNA 를 in vitro translation vector 에 cloning 된상태로제공받으실수있는 Gene Synthesis Service 도제공하고있습니다. 자세한내용은본책자 E. Gene Synthesis, 또는홈페이지 ( 를참조하시기바랍니다. 1) NH constructs (5 end 6x His-tagged template) T7 Promoter---RBS ATG 6x His-tag stop T7 terminator 2) CH constructs (3 end 6x His-tagged template) T7 Promoter---RBS ATG x His-tag - stop T7 terminator BIONEER 20

21 ExiProgen ProXpress PCR Template Kit Fast and Parallel Template DNA Preparation with PCR Cloning 과정없이 PCR 반응을이용하여무세포단백질발현 (cellfree protein expression) 용 template DNA 를제작할수있는 kit 로, 표적유전자증폭용 1 차 primer set 를제외한 template DNA 제조반응에필요한모든구성품 (2 차 primer set, PCR premix 등 ) 이 kit 에포함되어있습니다. 본 kit 로제작된단백질발현용 template DNA 는당사의무세포단백질발현및합성제품군에사용가능합니다. Rapid & Easy Preparation of Linear Template DNA (for Protein Expression) 목적단백질의유전자를포함하는 cdna, genomic DNA 등으로부터두단계의 PCR 반응을통해단백질합성용 template DNA를제조할수있으며, ExiProgen TM kit와연계하여하루만에목적단백질합성을완료할수있습니다. Accurate Protein Template by High Fidelity PCR 본 kit는당사의대표적인 long & high fidelity PCR premix인 AccuPower ProFi Taq PCR Premix를사용하여, PCR 반응시생길수있는 error rate를줄여주기때문에정확한 sequence를가진단백질발현용 template DNA를제작할수있습니다. High Efficiency 본 kit 를이용하여제조한단백질발현용 template 는 ExiProgen TM 에최적화되어소량의 DNA 만으로도 100 μg 이내의단백질을합성할수있습니다. (ex: template DNA 길이에따른최대단백질합성을위한권장 DNA 사용량 : 1 kb 이하 ng, 1 ~ 2 kb - 1 μg) Application 무세포단백질발현용 template DNA 제작에사용할수있습니다. Experimental Data st 1 PCR products M nd 2 PCR products M M2 Figure 1. Agarose gel data of template DNA synthesized by each PCR. Synthesis of linear template DNA with ExiProgen TM ProXpress PCR Template Kit. M1: 100 bp DNA Ladder (D-1030, Bioneer), M2: 1 kb DNA Ladder (D-1040, Bioneer) 1: SAV (Template - pt), 2: RNase H (Template - BL21 (DE3) gdna), 3: hgh (Template - pt) 4: CAT (Template - pbivt), 5: UDG (Template - BL21 (DE3) gdna), 6: AcGFP (Template - pbivt) 7: EVO (Template - pt), 8: RFP (Template - pivex), 9: Poly A polymerase (Template - pet15b) 21

22 ExiProgen ProXpress PCR Template Kit Figure 2. SDS-PAGE gel data of synthesized proteins. Each linear template DNA generated by ExiProgen TM ProXpress PCR Template Kit and those are used as template for protein synthesis with ExiProgen TM EC Protein Synthesis Kit. M: AccuLadder TM Protein Size Marker (Low) ( D-2020, Bioneer) 1: SAV (13 kda), 2: RNase H (20 kda), 3: hgh (23 kda), 4: CAT (26.5 kda), 5: UDG (28 kda) 6: AcGFP (28 kda), 7: EVO (30 kda), 8: RFP (31 kda), 9: Poly A polymerase (54 kda) Cat. no. Product Description K-7400 ExiProgen TM ProXpress PCR Template Kit, 16 rxns K-7401 ExiProgen TM ProXpress PCR Template Kit, 32 rxns Related Product N 차 primer F/R sets (N-terminus 6x His-tag), each 5 nmole 1 N 차 primer F/R sets (C-terminus 6x His-tag), each 5 nmole 1 K-2631 AccuPower ProFi Taq PCR PreMix, 96 tubes, 20 μl rxn/tube K-3034 AccuPrep PCR Purification Kit, 200 rxns K-3035 AccuPrep Gel Purification Kit, 200 rxns D-1030 AccuLadder TM 100 bp DNA Ladder, 250 μl (135 ng/μl) D kb DNA Ladder, 500 μl (130 ng/μl) 1 Oligo synthesis service 문의 BIONEER 22

23 ExiProgen Circular PCR Template Kit NEW Fast and Parallel Circular Template DNA Preparation with PCR Rapid & Easy Preparation of Circular Template DNA (for Protein Expression) 목적단백질의유전자를포함하는 cdna, genomic DNA 등으로부터세단계의 PCR 반응을통해단백질합성용 circular template DNA 를제조할수있으며, ExiProgen TM kit 와연계하여목적단백질합성을완료할수있습니다. Cloning 과정없이 PCR 을이용하여무세포단백질발현 (cell-free protein expression) 용 circular template DNA 를제작할수있는 kit 로, 표적유전자증폭용 1 차 primer set 를제외한 template DNA 제조반응에필요한모든구성품 (2 차 primer set, PCR premix 등 ) 이 kit 에포함되어있습니다. 본 kit 로제작된 circular DNA 는안정성이높아단백질발현시간이긴경우에도효율적으로단백질을발현할수있으며, 당사의무세포단백질발현및합성제품군에사용가능합니다. Template DNA Kit ExiProgen TM ProXpress PCR Template Kit ExiProgen TM Circular PCR Template Kit 사용가능한 Protein Synthesis Kit - ExiProgen TM EC Protein Synthesis Kit - ExiProgen TM EC Protein Synthesis Kit - ExiProgen TM EC-Maxi Protein Synthesis Kit - ExiProgen TM EC-Tagfree Protein Synthesis Kit - ExiProgen TM EC-Disulfide Protein Synthesis Kit Enhanced Stability PCR product 를 template 로사용할경우단백질이발현되는동안 nuclease 에의해 DNA 가분해되기때문에, 발현시간이많이요구되는 kit 를사용하게되면단백질효율이저하되는문제가있습니다. 본 kit 로부터합성되는 circular PCR template 는이러한문제점을극복하여목적단백질의발현효율을증대시켰습니다. Accurate Protein Template by High Fidelity PCR 본 kit 는당사의대표적인 long & high fidelity PCR premix 인 AccuPower ProFi Taq PCR PreMix 를사용하여, PCR 반응시생길수있는 error rate 를줄여주기때문에정확한 sequence 를가진단백질발현용 circular template DNA 를제작할수있습니다. High Efficiency 본 kit 를이용하여제조한단백질발현용 circular template 는 ExiProgen TM 에최적화되어소량의 DNA 만으로도단백질을합성할수있습니다. Application 무세포단백질발현용 circular template DNA 제작에사용될수있습니다. Experimental Data Figure 1. Agarose gel data of template circular DNA synthesized by three steps. Cirular PCR template is synthesized with ExiProgen TM Cirular PCR Template Kit. 23

24 ExiProgen Circular PCR Template Kit Figure 2. The performance test of ExiProgen TM Circular PCR Template Kit using ExiProgen TM EC-Maxi Protein Synthesis Kit M: protein marker 1: protein synthesis from 6 μg of plasmid DNA 2: protein synthesis from 0.3 μg of linear DNA 3: protein synthesis from 0.3 μg of circular DNA Figure 3. The performance test of ExiProgen TM Circular PCR Template Kit with various proteins using ExiProgen TM EC-Maxi Protein Synthesis Kit. (a) Linear DNA, (b) Circular DNA, (C) Plasmid DNA M: protein marker, 1: CAT (23 kda), 2: AcGFP (29 kda), 3: RFP (31 kda) Cat. no. Product Description K-7410 ExiProgen TM Circular PCR template Kit, 16 rxns K-7411 ExiProgen TM Circular PCR template Kit, 32 rxns Related Product N 차 primer F/R sets (N-terminus 6x His-tag), each 5 nmole 1 N 차 primer F/R sets (C-terminus 6x His-tag), each 5 nmole 1 K-2631 AccuPower ProFi Taq PCR PreMix, 96 tubes, 20 µl rxn/tube K-3034 AccuPrep PCR Purification Kit, 200 rxns K-3035 AccuPrep Gel Purification Kit, 200 rxns D-1030 AccuLadder TM 100 bp DNA Ladder, 250 µl (135 ng/ µl) D kb DNA Ladder, 500 µl (130 ng/µl) BIONEER 1 Oligo synthesis service 문의 24

25 pbivt Vector Set-1 바이오니아무세포단백질합성전용벡터 바이오니아의무세포단백질합성전용벡터인 pbivt 벡터세트 (pbivt-1, pbivt-2) 는당사의무세포단백질발현및합성의모든 kit 에적용가능합니다. 본벡터세트는 N-terminal (pbivt-1) 또는 C-terminal (pbivt-2) 에 6x His-tagging 이가능하며, 구조는아래와같습니다. Application 무세포단백질발현용 template DNA 제작에사용될수있습니다. Cat. no. Product Description K-7350 pbivt Vector Set-1 Related Product K-3030 AccuPrep Plasmid Extraction Kit, 200 rxns K-3034 AccuPrep PCR Purification Kit, 200 rxns D-1030 AccuLadder TM 100 bp DNA Ladder, 250 µl (135 ng/µl) D kb DNA Ladder, 500 μl (130 ng/μl) E-3061 T4 DNA Ligase, 20,000 U 25

26 04 Protein Service Protein Synthesis Service Standard Protein Synthesis Service 27 One-day Protein Synthesis Service 28 Gene to Protein Service Standard Gene to Protein Service 29 Cloning-free Gene to Protein Service 30

27 Standard Protein Synthesis Service Protein Synthesis Service 는원하는단백질을최소 1 주이내에공급받을수있는서비스입니다. 해당서비스는당사의전자동단백질합성장비 (ExiProgen TM ) 를이용하여단백질을합성해드리는서비스로, 다양한종류의단백질 ( 효소, 성장인자, hormone, 항원단백질, 항체등 ) 합성이가능합니다. 서비스진행방식 Protein Synthesis Service 는의뢰하시는 template DNA 의종류에따라서, 다음과같이두가지방식으로분류되어진행됩니다. Type I) E. coli expression vectors (ex. pet 등의 T7-based 발현벡터 ) Fast process E. coli 무세포발현법으로발현한단백질을 His-tag affinity 를이용하여정제하므로, 빠른시간내에정제된목적단백질을제공받으실수있습니다 (His-tag 이제거된상태의목적단백질도합성이가능합니다 ). Note) Plasmid DNA의전체크기에는제한이없으나, ORF (open reading frame) 를기준으로 0.3 kb~3.5 kb의시료만서비스가가능합니다. Type II) Non-expression vectors (ex. Cloning vector/ target gene) High performance 효소류, 성장인자, 호르몬, 항원단백질, 항체등다양한종류의단백질합성이가능하며, 또한세포배양방법으로생산이불가능했던독성단백질도합성이가능합니다. Compatibility 당사의단백질합성 kit 를이용하여단백질합성서비스가진행되므로, 필요한경우직접단백질을합성하실수도있습니다. Note) ORF (open reading frame) 를기준으로 0.3 kb~1.5 kb의시료만서비스가가능하며, 1.5 kb 이상의시료는발현벡터로구축한후, 의뢰하시기바랍니다. 시료제공량서비스비용서비스결과 3) 소요기간 4) 증량비용 5) E. coli expression vector 1) 10 μg 이상 (A260/280 : 1.7~2.0, A260/230 : >1.5) Non-expression vector 2) 2 μg 이상 600,000원 Note) 단백질합성이되지않을시, Data report 발송과함께 set up charge 100,000원이청구됩니다. 1. Data report Protein μg (0.1~1.0 mg/ml of solution form) 7 ~ 14일 100,000~300,000원 /100 μg 1) Plasmid DNA의양또는순도가조건에부합되지않는경우에는 DNA 증량서비스가추가되며, 추가비용이발생할수있습니다. 2) 일반 cloning용 vector 또는 PCR product로의뢰가가능하며, 이때에는 1차 primer sets 및 template DNA 제작서비스가추가되어, 추가비용 100,000 원이발생합니다. 3) 서비스비용은단백질 100 μg을합성해드리는비용이며, 이때합성량은목적단백질의순도와관계없이전체시료의양을의미합니다. 4) 소요기간은 working day 기준으로산정되며, DNA 시료상태또는단백질합성량에따라서증가할수있습니다. 5) 증량비용은단백질 100 μg 추가합성시에발생하는비용을의미하며, 합성난이도에따라서비용이차등부과됩니다. 27

28 One-day Protein Synthesis Service One-day Protein Synthesis Service 는당사의 ExiProgen TM EC Protein Synthesis Kit 를사용하여원하는단백질을합성해드리는서비스로, 합성결과를단하루만에받을수있습니다. 서비스진행방식 Fast process 단백질합성결과를단하루만에받을수있습니다. High performance 효소류, 성장인자, 호르몬, 항원단백질등다양한종류의단백질합성이가능하며, 또한세포배양방법으로생산이불가능했던독성단백질도합성이가능합니다. Compatibility 당사의단백질합성 kit (ExiProgen TM EC Protein Synthesis Kit) 를이용하여단백질합성서비스가진행되므로, 필요한경우직접단백질을합성하실수도있습니다. 시료제공량 E. coli expression vector 1) 10 μg 이상 (A260/280 : 1.7~2.0, A260/230 : >1.5) 서비스비용 300,000원 Note) 단백질합성이되지않을시, Data report 발송과함께 set up charge 100,000원이청구됩니다. 서비스결과 2) 1. Data report Protein - 0.1~1.0 mg/ml of solution 소요기간 3) 1일 1) Plasmid DNA의양또는순도가조건에부합되지않는경우에는 DNA 증량서비스가추가되며, 추가비용이발생할수있습니다. 2) Data report 및 1회반응에서합성된단백질을제공해드립니다. 추가적으로단백질이필요한경우에는, Standard Protein Synthesis Service 로전환됩니다. 3) 시료배송에소요되는기간은포함되지않습니다. BIONEER 28

29 Standard Gene to Protein Service Standard Gene to Protein Service 는당사의 Gene Synthesis Service 를기반으로합성된유전자를 E. coli 용 expression vector 에 cloning 한후, 단백질합성서비스가연계되어진행되는서비스입니다. 서비스진행방식 High performance 효소류, 성장인자, 호르몬, 항원단백질등다양한종류의단백질합성이가능하며, 또한세포배양방법으로생산이불가능했던독성단백질도합성이가능합니다. High quality Automatic DNA sequencer (ABI 3730) 를이용하여 100% sequence guarantee 의제품을제공받으실수있습니다. Codon optimization service KAIST 와공동개발한 codon optimization program 을이용하여 E. coli codon 에최적화된유전자를합성함으로서, 단백질발현효율을높일수있습니다. Compatibility 당사의단백질합성 kit 를이용하여단백질합성서비스가진행되므로, 필요한경우직접단백질을합성하실수도있습니다. 합성비용 1) 1,200,000원 (Normal sequence/300 bp~1 kb) 추가비용 2) Gene synthesis - 450원 /bp (1 kb 이상 ) 합성기간 1~1,500 bp 20~35 working days 1,501~3,000 bp 30~40 working days 시료제공 Gene Plasmid DNA (lysophilized form) ; 2~5 μg Protein 3) 100 μg (0.1~1.0 mg/ml) of solution 1) 주문후 5일이내취소시, gene 합성가의 50%, 5일이후취소시, gene 합성가의 80% 가청구됩니다. 2) High or low GC, repeat sequence, homo-polymeric runs 등이 sequence 내에포함되어있을경우, 합성단가및상향조정될수있습니다. 3) 단백질합성이되지않을시, Data report 발송과함께 set up charge 100,000원이부과됩니다. 단, 합성된유전자는정상제공되며, 유전자합성비용은청구됩니다. 29

30 Cloning-free Gene to Protein Service Cloning-free Gene to Protein Service 는당사의 AccuGeneBlock Service 를기반으로유전자를합성한후, PCR product 를이용하여단백질합성이진행되는서비스로, cloning 과정이필요없어, 보다빠른시간내에결과를받아보실수있습니다. 서비스소요시간비교 Fast process cloning, 세포배양과정없이 PCR product 를이용하여단백질합성이진행되는서비스로, 최소 2 주만에원하는유전자와단백질을받으실수있습니다. Codon optimization service KAIST 와공동개발한 codon optimization program 을이용하여 E. coli codon 에최적화된유전자를합성하므로, 단백질발현효율을높일수있습니다. Compatibility 당사의단백질합성 kit 를이용하여단백질합성서비스가진행되므로, 필요한경우직접단백질을합성하실수도있습니다. 합성비용합성기간 Gene 시료제공 Protein 1) 1,000,000원 (Normal sequence만서비스가능 /300 bp~1 kb) 10~15 working days PCR product (lyophilized form) ; 500 ng~1μg 100 μg (0.1~1.0 mg/ml) of solution 1) 단백질합성이되지않을시, Data report 발송과함께 set up charge 100,000원이부과됩니다. 단, 합성된유전자는정상제공되며, 유전자합성비용은청구됩니다. BIONEER 30