TH54566A3 - The process of separating the recombinant E protein obtained from E. To be used as an antibiotic protein in dengue diagnostic kits. - Google Patents
The process of separating the recombinant E protein obtained from E. To be used as an antibiotic protein in dengue diagnostic kits.Info
- Publication number
- TH54566A3 TH54566A3 TH9901000302A TH9901000302A TH54566A3 TH 54566 A3 TH54566 A3 TH 54566A3 TH 9901000302 A TH9901000302 A TH 9901000302A TH 9901000302 A TH9901000302 A TH 9901000302A TH 54566 A3 TH54566 A3 TH 54566A3
- Authority
- TH
- Thailand
- Prior art keywords
- protein
- recombinant
- coli
- separating
- react
- Prior art date
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract 11
- 102000004169 proteins and genes Human genes 0.000 title claims abstract 11
- 101710204837 Envelope small membrane protein Proteins 0.000 title claims abstract 5
- 238000000034 method Methods 0.000 title claims abstract 5
- 101710145006 Lysis protein Proteins 0.000 title claims abstract 3
- 208000001490 Dengue Diseases 0.000 title 1
- 206010012310 Dengue fever Diseases 0.000 title 1
- 238000009007 Diagnostic Kit Methods 0.000 title 1
- 239000003242 anti bacterial agent Substances 0.000 title 1
- 230000003115 biocidal effect Effects 0.000 title 1
- 208000025729 dengue disease Diseases 0.000 title 1
- 241000588724 Escherichia coli Species 0.000 claims abstract 5
- 239000011347 resin Substances 0.000 claims abstract 4
- 229920005989 resin Polymers 0.000 claims abstract 4
- 238000000746 purification Methods 0.000 claims abstract 3
- 102000004190 Enzymes Human genes 0.000 claims abstract 2
- 108090000790 Enzymes Proteins 0.000 claims abstract 2
- 101710088839 Replication initiation protein Proteins 0.000 claims abstract 2
- 229920004890 Triton X-100 Polymers 0.000 claims abstract 2
- 239000013504 Triton X-100 Substances 0.000 claims abstract 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims abstract 2
- 239000004202 carbamide Substances 0.000 claims abstract 2
- 238000003018 immunoassay Methods 0.000 claims abstract 2
- 210000003000 inclusion body Anatomy 0.000 claims abstract 2
- 238000002386 leaching Methods 0.000 claims abstract 2
- 238000000926 separation method Methods 0.000 claims abstract 2
- 239000010802 sludge Substances 0.000 claims abstract 2
- 238000001262 western blot Methods 0.000 claims abstract 2
Abstract
DC60 (30/06/42) กรรมวิธีการแยกโปรตีนรีคอมบิแนนต์ E จาก E. coli ซึ่งอยู่ในรูปของโปรตีนที่ไม่ละลายน้ำ หรือ inclusion bodies ซึ่งประกอบด้วยขั้นตอนที่สำคัญ 3 ส่วนคือ การแยกส่วนของตะกอนโปรตีนออกจากโปรตีนละลายน้ำอื่นๆ ของ E. coli ก่อน ตามด้วยการล้างตะกอนโปรตีนที่ได้ด้วยบัฟเฟอร์ที่มีส่วนประกอบของ Triton X-100 รวมทั้งขั้นตอนการล้างเร ซินหลังจากที่จับกับโปรตีน E ด้วยบัฟเฟอร์ที่มีส่วนประกอบของ 8M Urea pH 8 จำนวนอย่างน้อย 40 เท่าของปริมาตรเร ซิน ก่อนที่จะทำการชะโปรตีนออกจากเรซิน โดยกระบวนการทั้ง 3 ส่วนที่กล่าวมา จะทำให้ได้โปรตีนรีคอมบิแนนต์ E ที่มี ความบริสุทธิ์ สามารถทำปฏิกิริยาได้ดีกับ PCS และไม่ทำปฏิกิริยากับ PND ทั้งใน Western Blot Analysis และ Dot Enzyme Immunoassay (DEIA) DC60 (30/06/42) The method of separating recombinant E from E. coli, which is the insoluble protein or inclusion bodies, consists of three important steps: sludge separation. The protein was first removed from the other soluble proteins of E. coli, followed by a purification of the protein obtained with a buffer containing Triton X-100 as well as a purification procedure. Synthesin after binding to protein E with a buffer containing 8M Urea pH 8 at least 40 times the resin volume prior to leaching of the protein from the resin. By all 3 parts of the aforementioned processes Will yield the purified recombinant E protein It can react well with PCS and does not react with PND in both Western Blot Analysis and Dot Enzyme Immunoassay (DEIA).
Claims (1)
Publications (1)
| Publication Number | Publication Date |
|---|---|
| TH54566A3 true TH54566A3 (en) | 2002-12-24 |
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