Preparation and identification of monoclonal antibody against Homo sapiens hemoglobin alpha 2 (HBA2) / 中国实验血液学杂志
Journal of Experimental Hematology
;
(6): 823-826, 2007.
Article
in Chinese
| WPRIM
| ID: wpr-276814
ABSTRACT
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Hemoglobin A2
/
Molecular Sequence Data
/
Base Sequence
/
Alpha-Thalassemia
/
Bodily Secretions
/
Allergy and Immunology
/
Hybridomas
/
Mice, Inbred BALB C
/
Antibodies, Monoclonal
/
Antibody Specificity
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Journal of Experimental Hematology
Year:
2007
Type:
Article
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