The technical problems shoot for the production of anti low molecular weight hapten antibody.
Four kinds of small molecular weight haptens, Desmosine, Adriamycin, 4'-carboxycotinine and 5'-Fluorouracil were used as the immunizing antigens to obtain rabbits polyclonal antisera. All of these haptenes were conjugated to Bovine Serum Albumin (BSA) using 1-ethyl-3(3-dimethylaminopropyl)-carbodiimide (EDCl) reagent. The functional amino or/and carboxyl group in hapten molecules was utilized for conjugation with BSA by EDCl reagent. The hapten conjugated with BSA was mixed with Freund's Complete or Incomplete Adjuvant, and immunized to rabbits abdominal subcutaneous regions. More than 3 times immunized rabbit's antisera were evaluated the titer, and the specificity was investigated by ELISA method. The specific antisera against desmosine and adriamycin were produced. Anti-desmosine antibodies had no cross-reaction to molecular structure resemble pyridinoline. Anti-adriamycin antibodies reacted well to adriamycin and it could distinguish the differences of -epi type of adriamycin. The ELISA assay systems used here had such sensitivity. However the specificity possessing antisera against 4'-carboxycotinine and 5-Fluorouracil could not produce even though the antisera titers were elevated. The reasons why and what kinds of problems were there are discussed in this paper. For the production of the specific affinity possessing antisera, the length and distance between the carrier protein and the hapten and the shape of the functional groups were the important factors for production of the specific antisera which recognize free form hapten molecule.(ABSTRACT TRUNCATED AT 250 WORDS)
Watanabe
T
T
Department of Surgery, Tokai University School of Medicine, Japan.
Nakasaki
H
H
Kamijou
A
A
Mitomi
T
T
Matsuki
H
H
Kasuga
H
H
D000818:Animals; D000917:Antibody Formation; D003895:Desmosine / Q000276:immunology; D004317:Doxorubicin / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005472:Fluorouracil / Q000276:immunology; D006241:Haptens / Q000276:immunology*; D008297:Male; D008970:Molecular Weight; D011817:Rabbits
D006241:Haptens; D003895:Desmosine; D004317:Doxorubicin; D005472:Fluorouracil