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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.

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Authors+Show Affiliations

Bao F
College of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing 100044, China.
He J
No affiliation info available
Cao G
No affiliation info available
Yin F
No affiliation info available
Wang X
No affiliation info available
Pang S
No affiliation info available
Zhang Y
No affiliation info available
Hong T
No affiliation info available

MeSH

Amyloid beta-PeptidesAntibody SpecificityHumansPeptide FragmentsPeptide LibrarySingle-Chain Antibodies

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 -