[Preparation and characterization of monoclonal antibodies against the hemagglutinin of H9 subtype of avian influenza virus].
OBJECTIVE
To prepare monoclonal antibodies (mAb) against the hemagglutinin(HA) of H9 subtype of avian influenza virus (AIV).


METHODS
8 week-old female BALB/c mice were immunized with the inactivated vaccine of H9 subtype of AIV. Splenocytes from the immunized mice were fused with Sp2/0 myeloma cells, and positive hybridoma clones were screened by indirect ELISA and hemagglutination inhibition test (HI). The specificity of the mAb was characterized by ELISA, HI test, indirect immunofluorescence (IF) staining and Western blot.


RESULTS
Three hybridoma cell lines named 2H1, 2A3 and 1C8 against HA of AIV H9 were obtained. The HI titers of 3 mAbs were 1 x 2(8)-1 x 2(13), and the ELISA titers were 1 x 10(-7), 1 x 10(-5) and 5 x 10(-6), respectively. The immunoglobulin subclass of all 3 mAbs was IgG1. Western blot analysis confirmed that mAb 2H1 could recognize HA and reacted to 31 out of 32 isolates of H9 subtype of AIV.


CONCLUSIONS
Three mAbs recognizing HA of H9 subtype of AIV were obtained, which may provide an useful tool for the antigenic analysis, the serological diagnosis, the epidemiological survey and the evaluation of AIV vaccine.
Zeng
Yan-bing
YB
Key Laboratory of Animal Lemology, Ministry of Agriculture/College of Animal Science and Veterinary Medicine, Yangzhou University 225009, China.
Jiao
Xin-an
XA
Pan
Zhi-ming
ZM
Huang
Jin-lin
JL
Zhang
Ping-hu
PH
Zhang
Su-hua
SH
Sun
Quan-yun
QY
Liu
Xiu-fan
XF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology*; D005260:Female; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007074:Immunoglobulin G / Q000032:analysis; D009980:Influenza A virus / Q000145:classification / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007074:Immunoglobulin G; C109963:hemagglutinin, avian influenza A virus