[Development and characterization of monoclonal antibodies against H7 hemagglutinin of avian influenza virus].
OBJECTIVE
To prepare monoclonal antibodies (mAbs) against the hemagglutinin protein of H7 subtype of avian influenza virus (AIV).


METHODS
(6-8 weeks old) BALB/c mice of were immunized endermicly with H7 subtype of AIV. The splenocytes from the immunized mice were fused with Sp2/0-Ag-14 myeloma cells after the last immunization. Hybridoma cells were screened by hemagglutination (HA) and hemagglutination inhibition (HI) tests. The reactivity and specificity of mAbs were evaluated by HI test and Western blot assay.


RESULTS
Four hybridoma cell lines secreting specific mAbs named 2E2, 2A4, 5F5 and 7G5 were developed. The HI titers of these mAbs were 5 x 2(7)-5 x 2(11), and the immunoglobulin subclass of 2E2 was IgM, that of 2A4 was IgG1, and that of 5F5 and 7G5 was IgG2a. Western blot analysis confirmed that the mAbs only reacted with M(r) 75 000 HA protein of H7 subtype of AIV but did not react with the proteins of Newcastle disease virus (NDV). The results of HI reactivity assay suggested that 2E2, 5F5 and 7G5 only reacted with H7 subtype of AIV but did not react with other subtypes of AIV, NDV and infectious bronchitis virus (IBV). However 2A4 reacted not only with H7 subtype of AIV but also with H15N8 reference strain of AIV at low HI level.


CONCLUSIONS
These mAbs can be used as a useful tool to analyze the HA structure of AIV. They also provide the effective reagents for the rapid detection of H7 subtype of AIV.
Li
Yu-Jun
YJ
Key Laboratory of Animal Infectious Diseases of Ministry of Agriculture, Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, China.
Jiao
Xin-An
XA
Pan
Zhi-Ming
ZM
Sun
Lin
L
Wang
Chuan-Bin
CB
Zhang
Su-Hua
SH
Sun
Quan-Yun
QY
Liu
Xiu-Fan
XF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D006825:Hybridomas / Q000378:metabolism; D050440:Immunoglobulin Subunits / Q000276:immunology; D009980:Influenza A virus / Q000276:immunology* / Q000302:isolation & purification; D011200:Poultry / Q000821:virology*; D013997:Time Factors
D000911:Antibodies, Monoclonal; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D050440:Immunoglobulin Subunits