Antiserum immunogen

Antiserum Production The immunogen, carboxymethylmorphine-bovine-serum-albumin, is emulsified with equal volume of complete Freund s adjuvant. Initial immunization doses are injected into the New Zealand albino rabbits and later on this followed up with booster injections after a period of 6 weeks. The antiserum titer is determined with each booster dose injection and is duly harvested when the titre value is maximum. This is diluted suitably and employed in the radioimmunoassay. ... 

Immunization and Antibody Production The lypphilized immunogen obtained above is dissolved in normal saline and emulsified with equal volumes of complete Freund s adjuvant into a thick paste. Three New Zealand albino rabbits are immunized with the immunogen-paste through intradermal injections. The process is repeated twice at 2-weeks intervals followed by booster doses at monthly intervals. The antiserum is harvested when the plasma titer value is attained maximum. 

Antiserums produced in this manner are known as polyclonal because they contain many antibodies produced by different lymphocytes, each one responding either to different antigenic determinants of the original antigen or to other immunogenic substances in the injected material. Such a range of antibodies reduces the specificity of the method. 

The PAP method was pioneered by Sternberger in 1979 (1). The method uses an immunological sandwich amplification and the enzyme peroxidase to effect a signal. The unique feature of this procedure is the enzyme/antibody solution, the PAP immune complex. The horseradish peroxidase enzyme, itself an immunogenic protein, is used to inoculate a given species, and a polyclonal immune response is generated against the enzyme. This antiserum is harvested and placed in solution with the enzyme so that immune complexes form that... 

In addition Choi et al. utilized FlTC-labeled methamphetamine for competitive immunoassay of methamphetamine in urine (19). Instead of purified antibody or antibody fragment, antiserum was used. An aminobutyl derivative of metamphetamine was conjugated with proteins and used as an immunogen to produce antibodies for the assay. The free FITC-labeled tracer was well separated from the antibody-bound fraction, and the detection limit for the CE assay was lower than that for ELISA. 

The anti-S. faecalis antiserum reacted with Lac-BSA amd the glycan as shown in the bottom panels of Figure 8. The amti-lac antibodies from this serum also reacted with these two immunogens yielding a similar pattern as the unfractionated amtiserum. It should be noted that the amtiserum amd amtibody preparations exhibited partial identity with these two antigens. The anti-gal antibodies also reacted with Lac-BSA amd the glycan but the precipitin bamds were quite different. 

Two immunization procedures designed to enhance the immune response to multiple antigen mixtures have been reported recently. The cascade immunization technique (20) utilized in vitro depletion of E. coli proteins (ECPs) which had previously elicited an antibody response. The removal of these dominant immunogens from the mixture was accomplished by immunoabsorption with antibodies obtained from an earlier antiserum. The passive immunization procedure (21) relied on in vivo blocking of strong immunogens by the concurrent administration of early antiserum obtained previously. This latter report demonstrated the presence of an apparently poorly immunogenic ECP to which a humoral response could only be elicited by this passive procedure. 

This antiserum was used in the blind analysis of plasma samples from marijuana smokers. These samples were also analyzed by GC-MS at Battelle Institute. Regression analysis of the two sets of results showed good agreement RIA (ng/ml) = 1.2 GC + 0.14 with excellent correlation. Thus, the tritium labeled A8-THC combined with the antibody from the A9-immunogen is useful for the analysis of A9-THC in plasma. 

The immunisation procedure (at least 12 weeks with booster injections, addition of oil based adjuvants to stimulate the immunogenic response) gives rise to an antiserum representing a mixture of polyclonal antibodies which are produced by different immunocompetent cells and thus have different binding sites and affinities. 

Immunogen and Rabbit Antiserum Beraprost was linked to bovine serum albumin according to Erlanger B.F. et al. (1957). 

A partially purified Bacillus thurlnglensis var. israelensls (Bti) 6-endotoxin was used to Immunize rabbits. The antisera obtained have an improved specificity towards the mosquito larvacidal activity of the toxin, as opposed to antiserum raised when the whole crystal was used as immunogen. Using a two step/indirect ELISA (enzyme linked immunosorbent assay) procedure developed in our laboratory, fourteen experimental formulations were tested, and the results were compared with bioassays. An average of 69.1 international units 20% c.v. was found to associate with each ug of toxin detected by the ELISA. Our data indicate that when toxin specific antisera are available, Immunoassays can be used to predict the biological activity of Bti samples with reasonable accuracy. 

Immunoassay of Steroids.—Oestrone 3-methylphosphonothiolate, electrostatically complexed with methylated bovine serum albumen (BSA), is an effective immunogen for the production of an antiserum for oestrone 3-sulphate. The phosphorus-containing ester apparently mimics the size of the sulphate, but is more stable. Reductive amination, involving the coupling of 5a-androstane-3,17-dione and of 17/8-hydroxy-5a-androstan-3-one by reduction of their Schiff bases with NaBHsCN, resulted in coupling between the lysine residues of BSA and the C-3 position of the steroid, with up to 40 molecules of steroid per BSA molecule. These antigens led to effective antisera for the steroids. 

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