Antibodies Reactive with Specific Folic Acid Determinants<sup>*</sup>

Ricker, R; Stollar, B. David

doi:10.1021/bi00859a018

Cited by 26 publications

(2 citation statements)

References 5 publications

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“…After dialysis the dissolved and precipitated MenBGG (25-34 menadione residues/7S monomer BGG) were combined as a suspension. Folic acid2oBGG was prepared according to Ricker and Stollar, and the hapten is attached to the protein presumably by both its carboxyl and amino groups (12).…”

Section: Methodsmentioning

confidence: 99%

Antibodies with Multiple Binding Functions. Induction of Single Immunoglobin Species by Structurally Dissimilar Haptens

Várga

Konigsberg

Richards

1973

Proc. Natl. Acad. Sci. U.S.A.

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Anti-hapten sera prepared in rabbits contain individual immunoglobulin species capable of binding several pairs of structurally diverse haptens A and B. (A = inosine, uridine, menadione, vitamin K1, ribonuclease; B = 2,4-dinitrophenyl). In antisera against hapten A subjected to isoelectric focusing, there are many anti-A immunoglobulin species, but only a small, proportion of these bind both A and B. Immune sera characteristically show a high degree of specificity for the antigen used to elicit the immune response. It might be expected that the individual immunoglobulins that constitute the serum might also show a similar high degree of specificity. Talmage proposed a model whereby different immunoglobulins might each have a partial fit for antigen and in which the specificity of the immune serum was due to the sum of several different partial fits (1). The mouse immunoglobulin IgA2 myeloma protein 460 studied by Eisen (2) binds competitively the dissimilar haptens e-2,4-dinitrophenyl-ilysine (Dnp Lys) and 2-methyl-1,4-napthoquinone thioglycollate (MenTG) to different subsites in the combining region (3). Since several workers have obtained evidence suggesting that individual elicited antibodies may bind several structurally diverse antigens (2-6), we examined individual antibodies from anti-hapten sera prepared in rabbits to see if they also bind more than one structurally diverse antigen. We further asked that when one immunoglobulin binds both antigens A and B, whether both A and B induce the immune response of the same immunoglobulin. Isoelectric focusing and radioautography was done as described (8, 9), except that the polyacrylamide gels contained 4 M deionized urea. Antibodies were partially purified by precipitation with sodium sulfate (final concentration, 180 g/liter) three times and were dissolved in phosphate-buffered saline [0.01 M Na-phosphate-0.15 M NaCl (pH 7.4)] containing 4 M urea.Whole antibody populations were screened for crossreactivity (Varga, J. M. & Richards, F. F., in preparation). Briefly, partially hydrolyzed nylon mesh discs were used as initiator sites for polymerization of serine-N-carboxyanhydride (Pilot Chemicals) to polyserine chains. Antibodies against Dnp were coupled to the hydrophilic polyserine chains with glutaraldehyde. Crossreaction of a hapten with the antibodies against Dnp was measured by its inhibition of [8H ]eDnp-lysine binding.Dnp6oBGG and Tnp55BGG (2,4,6-trinitrophenyl-bovine gammaglobulin) were prepared according to Little and Eisen (10). U5&BGG (uridine58 BGG) was prepared by the cyanogen bromide activation method (11), and purified on a 3 X 60-cm Sephadex G-200 column developed with phosphatebuffered saline. U52KLH was prepared by a similar method.Men25-suBGG was prepared from 2-methyl-3-amino-1,4-naphthoquinone. Menadione bisulfate trihydrate (10 mmol) was dissolved in 10 ml of acetic acid. Sodium azide (10 mmol) dissolved in 5.0 ml of distilled water was added drop by drop over 20 min with stirring; 2-methyl-3-aminonaphthoquinone was precipitated, ...

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Section: Methodsmentioning

confidence: 99%

Antibodies with Multiple Binding Functions. Induction of Single Immunoglobin Species by Structurally Dissimilar Haptens

Várga

Konigsberg

Richards

1973

Proc. Natl. Acad. Sci. U.S.A.

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“…Since some of the coenzymes can compete successfully for the combining sites of the antibody, the effect of the purified antibodies on coenzyme-dependent enzymic activities can also be tested. Recently, antibodies specific for the coenzymes folic acid (Jaton and UngarWaron, 1967;Ricker and Stollar, 1967) and pyridoxal (Ungar-Waron and Sela, 1966;Cordoba et al, 1966) have been produced. Antibodies to pyridoxal did inhibit the pyridoxal-dependent enzyme, glutamic-oxalacetic transaminase (Ungar-Waron and Sela, 1966).…”

mentioning

confidence: 99%

Production of antibodies with specificity toward adenosine, uridine, and ribose 5'-phosphates

Vunakis¹,

Seaman²,

Setlow³

et al. 1968

Biochemistry

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During the photooxidation of guanine-containing nucleotides and dinucleotides, the guanine moiety is destroyed to yield a reactive photointermediate which condenses with the amino groups of a copolymer composed of glutamyl and lysyl residues. Immunization of rabbits with such conjugates complexed to methylated bovine serum albumin results in the production of antibodies directed toward the haptens as well as toward the random copolymer. The specificities of the antisera to conjugates obtained with guanosine 5'-phosphate (GMP-5'), GpA, and GpU photoproducts were examined by complement fixation. The antibodies directed toward the mononucleotide coniugate show specificity toward the ribose 5 '-phosphate moieties of the nucleotides. In sera of rabbits immunized with photooxidized GpA and GpU conjugates, the antibodies are directed toward the 5 '-nucleotides, adenosine 5'-phosphate (AMP-5') and uridine 5 '-phos-

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Principles of synthesis of conjugated antigens

Polevaya

Ковалев

1978

Pharm Chem J

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Antibodies Reactive with Specific Folic Acid Determinants^*

Cited by 26 publications

References 5 publications

Antibodies with Multiple Binding Functions. Induction of Single Immunoglobin Species by Structurally Dissimilar Haptens

Antibodies with Multiple Binding Functions. Induction of Single Immunoglobin Species by Structurally Dissimilar Haptens

Production of antibodies with specificity toward adenosine, uridine, and ribose 5'-phosphates

Principles of synthesis of conjugated antigens

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Antibodies Reactive with Specific Folic Acid Determinants*

Cited by 26 publications

References 5 publications

Antibodies with Multiple Binding Functions. Induction of Single Immunoglobin Species by Structurally Dissimilar Haptens

Antibodies with Multiple Binding Functions. Induction of Single Immunoglobin Species by Structurally Dissimilar Haptens

Production of antibodies with specificity toward adenosine, uridine, and ribose 5'-phosphates

Principles of synthesis of conjugated antigens

Contact Info

Product

Resources

About

Antibodies Reactive with Specific Folic Acid Determinants^*