Shorter development of immunoassay for drugs: application of the novel RIMMS technique enables rapid production of monoclonal antibodies to ranitidine.
High affinity, specific murine monoclonal antibodies have been produced for ranitidine using the novel RIMMS (repetitive immunizations, multiple sites) technique. We demonstrate that this technique can be employed to produce high affinity monoclonal antibodies to drug haptens in approximately 1 month; whereas, conventional techniques typically require 3-9 months. Polyclonal antiserum development typically requires at least 6 months. Consequently, RIMMS has a clear impact allowing reagent antibodies to be available earlier in the drug development process. Isotyping studies demonstrated that the developed antibodies are either IgG1 or IgG2b immunoglobulins which confirms that the technique produces class-switched, affinity matured reagent antibodies. The most promising monoclonal antibody for quantitative applications afforded similar sensitivity, by competitive ELISA, to the established sheep polyclonal anti-ranitidine sera. The calibration range, estimated as the limits between the asymptotic regions of calibration graphs, is 0.5-41.2 ng ranitidine per well. Specificity studies indicated that the monoclonal antibody afforded superior selectivity, yielding only 4.1% cross-reactivity with the ranitidine sulphoxide metabolite; the corresponding value for the antiserum was 8.6%. Both reagents had similar cross-reactivities with the N-oxide metabolite.
Wring
S A
SA
Bioanalysis and Drug Metabolism Division, Glaxo Wellcome Research Inc., Research Triangle Park, NC 27709, USA. saw22503@glaxowellcom.com
Kilpatrick
K E
KE
Hutchins
J T
JT
Witherspoon
S M
SM
Ellis
B
B
Jenner
W N
WN
Serabjit-Singh
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002138:Calibration; D002459:Cell Fusion; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006241:Haptens / Q000276:immunology; D006825:Hybridomas / Q000276:immunology; D007115:Immunization Schedule; D007202:Indicators and Reagents; D051379:Mice; D011506:Proteins / Q000737:chemistry / Q000276:immunology; D011863:Radioimmunoassay; D011899:Ranitidine / Q000737:chemistry / Q000276:immunology*; D012996:Solutions
D000911:Antibodies, Monoclonal; D006241:Haptens; D007202:Indicators and Reagents; D011506:Proteins; D012996:Solutions; D011899:Ranitidine