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[Isolation and identification of a human single chain Fv antibody against amyloid-beta 1-42 soluble oligomers from a human phage display library].
Sheng Wu Gong Cheng Xue Bao. 2009 Aug; 25(8):1195-203.SW

Abstract

To get specific scFv (Single-chain fragment variable) antibody against soluble Abeta1-42(Amyloid-beta) oligomers, we constructed a human single-chain Fv (scFv) antibody library by phage display technology. Using RT-PCR, we amplified the variable heavy (VH) and variable light (VL) genes from peripheral blood lymphocytes (PBL). Then we obtained the scFv fragments through SOE-PCR, and the scFv fragments were cloned into the vector pCANTAB5E and electroporated into competent Escherichia coli TG1 cells. Consequently, a scFv phage display library containing 2.5 x 10(9) clones was constructed. The recombinant phagemids were rescued by reinfection of helper phage M13K07. Recombinant phages specific for Abeta1-42 oligomers were enriched after four rounds of biopanning and the antigen-positive clones were selected from the enriched clones by phage ELISA. Positive clone B19 was used to infect E. coli HB2151 to express soluble scFv antibody. SDS-PAGE and Western blotting analysis showed that the soluble scFv B19 antibody was expressed successfully and could bind specifically to Abeta1-42 trimer and protofiber. The specific scFv against Abeta1-42 oligomers can be used in the therapeutic research on Alzheimer's disease.

Authors+Show Affiliations

College of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing 100044, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

19938457

Citation

Bao, Fuxiang, et al. "[Isolation and Identification of a Human Single Chain Fv Antibody Against Amyloid-beta 1-42 Soluble Oligomers From a Human Phage Display Library]." Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, vol. 25, no. 8, 2009, pp. 1195-203.
Bao F, He J, Cao G, et al. [Isolation and identification of a human single chain Fv antibody against amyloid-beta 1-42 soluble oligomers from a human phage display library]. Sheng Wu Gong Cheng Xue Bao. 2009;25(8):1195-203.
Bao, F., He, J., Cao, G., Yin, F., Wang, X., Pang, S., Zhang, Y., & Hong, T. (2009). [Isolation and identification of a human single chain Fv antibody against amyloid-beta 1-42 soluble oligomers from a human phage display library]. Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, 25(8), 1195-203.
Bao F, et al. [Isolation and Identification of a Human Single Chain Fv Antibody Against Amyloid-beta 1-42 Soluble Oligomers From a Human Phage Display Library]. Sheng Wu Gong Cheng Xue Bao. 2009;25(8):1195-203. PubMed PMID: 19938457.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Isolation and identification of a human single chain Fv antibody against amyloid-beta 1-42 soluble oligomers from a human phage display library]. AU - Bao,Fuxiang, AU - He,Jinsheng, AU - Cao,Guifang, AU - Yin,Fan, AU - Wang,Xin, AU - Pang,Siyuan, AU - Zhang,Ying, AU - Hong,Tao, PY - 2009/11/27/entrez PY - 2009/11/27/pubmed PY - 2011/4/29/medline SP - 1195 EP - 203 JF - Sheng wu gong cheng xue bao = Chinese journal of biotechnology JO - Sheng Wu Gong Cheng Xue Bao VL - 25 IS - 8 N2 - To get specific scFv (Single-chain fragment variable) antibody against soluble Abeta1-42(Amyloid-beta) oligomers, we constructed a human single-chain Fv (scFv) antibody library by phage display technology. Using RT-PCR, we amplified the variable heavy (VH) and variable light (VL) genes from peripheral blood lymphocytes (PBL). Then we obtained the scFv fragments through SOE-PCR, and the scFv fragments were cloned into the vector pCANTAB5E and electroporated into competent Escherichia coli TG1 cells. Consequently, a scFv phage display library containing 2.5 x 10(9) clones was constructed. The recombinant phagemids were rescued by reinfection of helper phage M13K07. Recombinant phages specific for Abeta1-42 oligomers were enriched after four rounds of biopanning and the antigen-positive clones were selected from the enriched clones by phage ELISA. Positive clone B19 was used to infect E. coli HB2151 to express soluble scFv antibody. SDS-PAGE and Western blotting analysis showed that the soluble scFv B19 antibody was expressed successfully and could bind specifically to Abeta1-42 trimer and protofiber. The specific scFv against Abeta1-42 oligomers can be used in the therapeutic research on Alzheimer's disease. SN - 1000-3061 UR - https://www.unboundmedicine.com/medline/citation/19938457/[Isolation_and_identification_of_a_human_single_chain_Fv_antibody_against_amyloid_beta_1_42_soluble_oligomers_from_a_human_phage_display_library]_ L2 - https://www.lens.org/lens/search/patent/list?q=citation_id:19938457 DB - PRIME DP - Unbound Medicine ER -