PMID- 22695166
OWN - NLM
STAT- MEDLINE
DCOM- 20121206
LR  - 20211021
IS  - 1556-679X (Electronic)
IS  - 1556-6811 (Print)
IS  - 1556-679X (Linking)
VI  - 19
IP  - 8
DP  - 2012 Aug
TI  - Enhanced bovine herpesvirus type 1 neutralization by multimerized single-chain 
      variable antibody fragments regardless of differential glycosylation.
PG  - 1150-7
LID - 10.1128/CVI.00130-12 [doi]
AB  - Single-chain variable antibody fragments (scFvs) with a 2-amino-acid linker 
      capable of multimerization as di-, tri-, or tetrabodies that neutralize bovine 
      herpesvirus type 1 (BoHV-1) in vitro were constructed and expressed in Pichia 
      pastoris. In contrast to the monomeric form, multimeric scFvs had a higher virus 
      neutralization potency, as evidenced by a 2-fold increase in their ability to 
      neutralize BoHV-1 due to avidity effects. Mass spectrum (quadrupole time of 
      flight [Q-TOF]) analyses of multimeric scFv demonstrated extensive heterogeneity 
      due to differential cleavage, variable glycosylation (1 to 9 mannose residues), 
      and the incorporation of minor unidentified adducts. Regardless of the 
      differential glycosylation patterns, the scFvs recognized non-gB or -gE target 
      viral epitopes in the BoHV-1 envelope fraction in a Western blot and also 
      neutralized BoHV-1 in infected Madin-Darby kidney (MDBK) cells in vitro. Indirect 
      evidence for the noncovalent multimerization of scFv was the presence of a major 
      peak of multimerized scFv without a His tag (due to differential cleavage) in the 
      Q-TOF profile, unlike monomeric scFv, which copurified with normally His-tagged 
      scFv and recognized the target antigen. Overall, differentially glycosylated 
      recombinant scFvs against BoHV-1 with a short linker (2 amino acids) are capable 
      of assembly into functional multimers that confer high avidity, resulting in 
      increased virus neutralization in vitro compared to that of monovalent scFv with 
      a long (18-amino-acid) flexible linker. Overall, recombinant multimerized 
      scFv5-2L potentially provides a high-potency therapeutic and immunodiagnostic 
      reagent against BoHV-1, which is suitable for passive immunization and topical 
      application.
FAU - Pasman, Yfke
AU  - Pasman Y
AD  - Department of Molecular and Cellular Biology, University of Guelph, Guelph, 
      Ontario, Canada.
FAU - Nagy, Eva
AU  - Nagy E
FAU - Kaushik, Azad K
AU  - Kaushik AK
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120613
PL  - United States
TA  - Clin Vaccine Immunol
JT  - Clinical and vaccine immunology : CVI
JID - 101252125
RN  - 0 (Antibodies, Neutralizing)
RN  - 0 (Antibodies, Viral)
RN  - 0 (Epitopes)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Single-Chain Antibodies)
SB  - IM
MH  - Antibodies, Neutralizing/*immunology
MH  - Antibodies, Viral/*immunology
MH  - Base Sequence
MH  - Blotting, Western
MH  - Epitopes/immunology
MH  - Gene Expression
MH  - Glycosylation
MH  - Herpesvirus 1, Bovine/*immunology
MH  - Mass Spectrometry
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - Neutralization Tests
MH  - Pichia/genetics
MH  - Protein Multimerization
MH  - Recombinant Proteins/chemistry/genetics/immunology/metabolism
MH  - Single-Chain Antibodies/chemistry/genetics/*immunology/metabolism
PMC - PMC3416088
EDAT- 2012/06/15 06:00
MHDA- 2012/12/10 06:00
PMCR- 2013/02/01
CRDT- 2012/06/15 06:00
PHST- 2012/06/15 06:00 [entrez]
PHST- 2012/06/15 06:00 [pubmed]
PHST- 2012/12/10 06:00 [medline]
PHST- 2013/02/01 00:00 [pmc-release]
AID - CVI.00130-12 [pii]
AID - 00130-12 [pii]
AID - 10.1128/CVI.00130-12 [doi]
PST - ppublish
SO  - Clin Vaccine Immunol. 2012 Aug;19(8):1150-7. doi: 10.1128/CVI.00130-12. Epub 2012 
      Jun 13.