Neutralisation and binding of VHS virus by monovalent antibody fragments

P. M. Cupit, N. Lorenzen, Gillian Louise Strachan, G. J. L. Kemp, Christopher John Secombes, C. Cunningham

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We have previously reported the cloning and characterisation of the heavy and light chain variable domain genes encoding three monoclonal antibodies (Mabs) that bind viral haemorrhagic septicaemia virus (VHSV). Two of these antibodies, 3F1H10 and 3F1A2 both neutralised the virus though 3F1A2 appeared to recognise a broader range of virus isolates. The variable domains of these two antibodies differ by only four residues (Lorenzen et al., 2000a. Fish Shellfish Immunol. 10, 129-142). To further study the mechanism of neutralisation, Fab fragments as well as a series of recombinant bacterial single chain antibody (scAb) fragments were generated from the three anti-VHSV Mabs and their variable domain genes, respectively. Fabs and scAbs derived from the neutralising Mabs were both able to neutralise the VHSV type 1 isolate DK-F1. In addition, a series of scAb fragments were produced using the 3F1H10 variable heavy (VH) chain and variable light (V kappa) chain domains but containing, either alone or in dual combination, each of the four different residues present in 3F1A2. The dissociation constants of Mabs 3F1H10 and 3F1A2 and their respective Fab and scAb fragments were measured by BIAcore analysis and found to correlate with the capacity of each molecule to neutralise DK-F1. These investigations, together with computer assisted molecular analysis of the theoretical influence of each mutation on antigen binding, led to the identification of a single mutation at position 35a in the VH domain as having the most marked impact on viral neutralisation. (C) 2001 Published by Elsevier Science B.V.

Original languageEnglish
Pages (from-to)47-56
Number of pages9
JournalVirus Research
Volume81
DOIs
Publication statusPublished - 2001

Keywords

  • egtved virus
  • viral haemorrhagic septicaemia virus
  • single chain antibody
  • virus neutralisation
  • MONOCLONAL-ANTIBODIES
  • RABIES VIRUS
  • PROTEIN
  • GLYCOPROTEIN
  • FISH

Cite this

Cupit, P. M., Lorenzen, N., Strachan, G. L., Kemp, G. J. L., Secombes, C. J., & Cunningham, C. (2001). Neutralisation and binding of VHS virus by monovalent antibody fragments. Virus Research, 81, 47-56. https://doi.org/10.1016/S0168-1702(01)00354-9

Neutralisation and binding of VHS virus by monovalent antibody fragments. / Cupit, P. M.; Lorenzen, N.; Strachan, Gillian Louise; Kemp, G. J. L.; Secombes, Christopher John; Cunningham, C.

In: Virus Research, Vol. 81, 2001, p. 47-56.

Research output: Contribution to journalArticle

Cupit, P. M. ; Lorenzen, N. ; Strachan, Gillian Louise ; Kemp, G. J. L. ; Secombes, Christopher John ; Cunningham, C. / Neutralisation and binding of VHS virus by monovalent antibody fragments. In: Virus Research. 2001 ; Vol. 81. pp. 47-56.
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AU - Lorenzen, N.

AU - Strachan, Gillian Louise

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AU - Secombes, Christopher John

AU - Cunningham, C.

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AB - We have previously reported the cloning and characterisation of the heavy and light chain variable domain genes encoding three monoclonal antibodies (Mabs) that bind viral haemorrhagic septicaemia virus (VHSV). Two of these antibodies, 3F1H10 and 3F1A2 both neutralised the virus though 3F1A2 appeared to recognise a broader range of virus isolates. The variable domains of these two antibodies differ by only four residues (Lorenzen et al., 2000a. Fish Shellfish Immunol. 10, 129-142). To further study the mechanism of neutralisation, Fab fragments as well as a series of recombinant bacterial single chain antibody (scAb) fragments were generated from the three anti-VHSV Mabs and their variable domain genes, respectively. Fabs and scAbs derived from the neutralising Mabs were both able to neutralise the VHSV type 1 isolate DK-F1. In addition, a series of scAb fragments were produced using the 3F1H10 variable heavy (VH) chain and variable light (V kappa) chain domains but containing, either alone or in dual combination, each of the four different residues present in 3F1A2. The dissociation constants of Mabs 3F1H10 and 3F1A2 and their respective Fab and scAb fragments were measured by BIAcore analysis and found to correlate with the capacity of each molecule to neutralise DK-F1. These investigations, together with computer assisted molecular analysis of the theoretical influence of each mutation on antigen binding, led to the identification of a single mutation at position 35a in the VH domain as having the most marked impact on viral neutralisation. (C) 2001 Published by Elsevier Science B.V.

KW - egtved virus

KW - viral haemorrhagic septicaemia virus

KW - single chain antibody

KW - virus neutralisation

KW - MONOCLONAL-ANTIBODIES

KW - RABIES VIRUS

KW - PROTEIN

KW - GLYCOPROTEIN

KW - FISH

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VL - 81

SP - 47

EP - 56

JO - Virus Research

JF - Virus Research

SN - 0168-1702

ER -