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Generation of chicken single chain antibody variable fragments (scFv) that differentiate and neutralize infectious bursal disease virus (IBDV).
Arch Virol. 2003 Mar; 148(3):497-515.AV

Abstract

Phage-displayed recombinant antibody libraries derived from splenic mRNA of chickens immunized with an Australian strain of infectious bursal disease virus (IBDV) were constructed as single chain variable fragments (scFv) by either overlap extension polymerase chain reaction (PCR) or sequential ligation of the individual heavy (V(H)) and light (V(L)) chain variable gene segments. Sequential cloning of the individual V(H) and V(L) genes into a newly constructed pCANTAB-link vector containing the synthetic linker sequence (Gly(4)Ser)(3) was more efficient than cloning by overlap extension PCR, increasing the library size 500 fold. Eighteen IBDV specific antibodies with unique scFv sequences were identified after panning the library against the immunizing antigen. Eight of the clones contained an identical V(H) gene but unique V(L) genes. In ELISA analysis using a panel of Australian and overseas IBDV strains, one scFv antibody was able to detect all strains, whilst 3 others could discriminate between Australian and overseas strains, classical and variant strains and Australian field strains and vaccine strains. In addition, some scFvs showed significant neutralization titres in vitro. This report shows that generation of chicken antibodies in vitro by recombinant means has considerable potential for producing antibodies of diverse specificity and neutralizing capacity.

Authors+Show Affiliations

CSIRO Livestock Industries, Australian Animal Health Laboratory, Geelong, VIC, Australia. sandra.sapats@csiro.auNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

12607101

Citation

Sapats, S I., et al. "Generation of Chicken Single Chain Antibody Variable Fragments (scFv) That Differentiate and Neutralize Infectious Bursal Disease Virus (IBDV)." Archives of Virology, vol. 148, no. 3, 2003, pp. 497-515.
Sapats SI, Heine HG, Trinidad L, et al. Generation of chicken single chain antibody variable fragments (scFv) that differentiate and neutralize infectious bursal disease virus (IBDV). Arch Virol. 2003;148(3):497-515.
Sapats, S. I., Heine, H. G., Trinidad, L., Gould, G. J., Foord, A. J., Doolan, S. G., Prowse, S., & Ignjatovic, J. (2003). Generation of chicken single chain antibody variable fragments (scFv) that differentiate and neutralize infectious bursal disease virus (IBDV). Archives of Virology, 148(3), 497-515.
Sapats SI, et al. Generation of Chicken Single Chain Antibody Variable Fragments (scFv) That Differentiate and Neutralize Infectious Bursal Disease Virus (IBDV). Arch Virol. 2003;148(3):497-515. PubMed PMID: 12607101.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Generation of chicken single chain antibody variable fragments (scFv) that differentiate and neutralize infectious bursal disease virus (IBDV). AU - Sapats,S I, AU - Heine,H G, AU - Trinidad,L, AU - Gould,G J, AU - Foord,A J, AU - Doolan,S G, AU - Prowse,S, AU - Ignjatovic,J, PY - 2003/2/28/pubmed PY - 2003/5/29/medline PY - 2003/2/28/entrez SP - 497 EP - 515 JF - Archives of virology JO - Arch Virol VL - 148 IS - 3 N2 - Phage-displayed recombinant antibody libraries derived from splenic mRNA of chickens immunized with an Australian strain of infectious bursal disease virus (IBDV) were constructed as single chain variable fragments (scFv) by either overlap extension polymerase chain reaction (PCR) or sequential ligation of the individual heavy (V(H)) and light (V(L)) chain variable gene segments. Sequential cloning of the individual V(H) and V(L) genes into a newly constructed pCANTAB-link vector containing the synthetic linker sequence (Gly(4)Ser)(3) was more efficient than cloning by overlap extension PCR, increasing the library size 500 fold. Eighteen IBDV specific antibodies with unique scFv sequences were identified after panning the library against the immunizing antigen. Eight of the clones contained an identical V(H) gene but unique V(L) genes. In ELISA analysis using a panel of Australian and overseas IBDV strains, one scFv antibody was able to detect all strains, whilst 3 others could discriminate between Australian and overseas strains, classical and variant strains and Australian field strains and vaccine strains. In addition, some scFvs showed significant neutralization titres in vitro. This report shows that generation of chicken antibodies in vitro by recombinant means has considerable potential for producing antibodies of diverse specificity and neutralizing capacity. SN - 0304-8608 UR - https://www.unboundmedicine.com/medline/citation/12607101/Generation_of_chicken_single_chain_antibody_variable_fragments__scFv__that_differentiate_and_neutralize_infectious_bursal_disease_virus__IBDV__ L2 - https://dx.doi.org/10.1007/s00705-002-0931-2 DB - PRIME DP - Unbound Medicine ER -