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Functional recombinant single-chain variable fragment antibody against Agkistrodon acutus venom.
Exp Ther Med. 2019 May; 17(5):3768-3774.ET

Abstract

Agkistrodon acutus bites are conventionally treated with animal-derived antivenom, the use of which is limited due to allergic reactions and serum sickness. Thus in the present study, the genes of humanized antibodies produced in response to A. acutus venom were extracted from lymphocytes from patients bitten by A. acutus. A single-chain variable fragment (scFv) library against venom was constructed using a T7 phage display system. ScFv genes that exhibited high affinity to venom were selected by library biopanning. An expression system was constructed for antivenom scFv fused with 6×His tag at its N- and C-terminus using pET-28a (+) vector. The scFv proteins could achieve functional and soluble expression in Escherichia coli via the auto-induction method. The purity and activity of the scFv genes and proteins were confirmed by SDS-PAGE, western blotting and ELISA. The results indicated that three soluble scFv proteins exhibited specific affinity to A. acutus venom and were harvested via the auto-induction method.

Authors+Show Affiliations

Department of Emergency, Southwest Hospital of the Third Military Medical University, Chongqing 400038, P.R. China.Department of Emergency, Southwest Hospital of the Third Military Medical University, Chongqing 400038, P.R. China.Department of Emergency, Southwest Hospital of the Third Military Medical University, Chongqing 400038, P.R. China.Department of Emergency, Southwest Hospital of the Third Military Medical University, Chongqing 400038, P.R. China.Department of Emergency, Southwest Hospital of the Third Military Medical University, Chongqing 400038, P.R. China.Department of Emergency, Southwest Hospital of the Third Military Medical University, Chongqing 400038, P.R. China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30988762

Citation

Zhang, Lei, et al. "Functional Recombinant Single-chain Variable Fragment Antibody Against Agkistrodon Acutus Venom." Experimental and Therapeutic Medicine, vol. 17, no. 5, 2019, pp. 3768-3774.
Zhang L, Cao Y, Liu M, et al. Functional recombinant single-chain variable fragment antibody against Agkistrodon acutus venom. Exp Ther Med. 2019;17(5):3768-3774.
Zhang, L., Cao, Y., Liu, M., Chen, X., Xiang, Q., & Tian, J. (2019). Functional recombinant single-chain variable fragment antibody against Agkistrodon acutus venom. Experimental and Therapeutic Medicine, 17(5), 3768-3774. https://doi.org/10.3892/etm.2019.7348
Zhang L, et al. Functional Recombinant Single-chain Variable Fragment Antibody Against Agkistrodon Acutus Venom. Exp Ther Med. 2019;17(5):3768-3774. PubMed PMID: 30988762.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Functional recombinant single-chain variable fragment antibody against Agkistrodon acutus venom. AU - Zhang,Lei, AU - Cao,Yuliang, AU - Liu,Minghua, AU - Chen,Xiangyu, AU - Xiang,Qiang, AU - Tian,Jun, Y1 - 2019/03/06/ PY - 2016/12/26/received PY - 2018/04/11/accepted PY - 2019/4/17/entrez PY - 2019/4/17/pubmed PY - 2019/4/17/medline KW - Agkistrodon acutus venom KW - auto-induction KW - phage display KW - single-chain variable fragment SP - 3768 EP - 3774 JF - Experimental and therapeutic medicine JO - Exp Ther Med VL - 17 IS - 5 N2 - Agkistrodon acutus bites are conventionally treated with animal-derived antivenom, the use of which is limited due to allergic reactions and serum sickness. Thus in the present study, the genes of humanized antibodies produced in response to A. acutus venom were extracted from lymphocytes from patients bitten by A. acutus. A single-chain variable fragment (scFv) library against venom was constructed using a T7 phage display system. ScFv genes that exhibited high affinity to venom were selected by library biopanning. An expression system was constructed for antivenom scFv fused with 6×His tag at its N- and C-terminus using pET-28a (+) vector. The scFv proteins could achieve functional and soluble expression in Escherichia coli via the auto-induction method. The purity and activity of the scFv genes and proteins were confirmed by SDS-PAGE, western blotting and ELISA. The results indicated that three soluble scFv proteins exhibited specific affinity to A. acutus venom and were harvested via the auto-induction method. SN - 1792-0981 UR - https://www.unboundmedicine.com/medline/citation/30988762/Functional_recombinant_single-chain_variable_fragment_antibody_against_Agkistrodon_acutus_venom L2 - https://www.ingentaconnect.com/openurl?genre=article&issn=1792-0981&volume=17&issue=5&spage=3768&aulast=Zhang DB - PRIME DP - Unbound Medicine ER -
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