[Preparation and identification of monoclonal antibody against Homo sapiens hemoglobin alpha 2 (HBA2)].
This study was purposed to prepare and identify monoclonal antibodies (McAbs) against Homo sapiens hemoglobin alpha 2 (HBA2). Normal human fetal liver tissues were homogenized, and human liver nuclear proteins were isolated by centrifugation. The total human fetal liver nuclear proteins were used to immunize BALB/c mice for preparing McAbs by hybridoma technique. The McAbs specificity was identified by ELISA, Western blot, and immunohistochemistry. The antigen was identified by Uni-ZAP expression library screening. The results showed that one hybridoma cell line, AEE091, secreting specific McAb against HBA2 was established. The Ig subclass of this McAb was IgG1 (kappa). Data from immunohistochemistry assay showed that AEE091 could recognize human liver nuclear protein. Using AEE091 McAb, isolation of the protein antigen by IP revealed that AEE091 McAb could recognize 15 kD protein. Screening the Uni-ZAP XR pre-made liver cDNA library with AEE091 hybridoma cell supernatants demonstrated that AEE091 McAb specially reacted with HBA2. It is concluded that a hybridoma cell line stably secreting specific McAb against HBA2 is established. The specific McAb against HBA2 would be very useful for studying HBA2 function and screening thalassemia.
Chen
Zhi-Cheng
ZC
Department of Immunology, Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China.
Yang
Jin-Ju
JJ
Liu
Rong
R
Qu
Hai-Xia
HX
Wang
Wan
W
Liu
Li
L
Liu
Xiao-Lan
XL
Chen
Yong
Y
Liu
Ying
Y
Gao
Jian-En
JE
Sun
Qi-Hong
QH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D001483:Base Sequence; D006443:Hemoglobin A2 / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D017085:alpha-Thalassemia / Q000276:immunology
D000911:Antibodies, Monoclonal; D006443:Hemoglobin A2