Abstract
Method of Immunization
Goose immunoglobulin purified from serum.
Method of Immunization
50 μg purified goose Ig emulsions in Freund's complete adjuvant were immunized with subcutaneous injections in BALB/c (6∼8 weeks old) mice and boosted with an additional 50 μg/mouse of goose immunoglobulin intraperitoneally without adjuvant on day 21.
Parental Cell Line Used for Fusion
Spleen cells from immunized BALB/c mice and mouse myeloma cell line SP2/0.
Selection and Cloning Procedure
The positive clone was selected and cloned three times using indirect ELISA in which plates were coated with recombinant protein rGoCL.(1,2)
Heavy and Light Chains of Immunoglobulin
Heavy chain, γ1; Light chain, κ.
Specificity
The specificity was determined using Western blot, indirect ELISA, and flow cytometry. The results indicated that MAb 1B11 only reacted with the light chain of goose immunoglobulin, but showed no cross-reaction with Gallus gallus, G. domesticus, turkey, duck, cow, sheep, pig, rabbit, Anas platyrhynchos, and Grus japonensis.
Specific Antigen Identified
The light chain of goose immunoglobulin from serum and bile, as well as the peripheral blood B lymphocytes were used as specific antigen to identify MAb 1B11.