Production and characterization of antibodies cross-reactive with major aflatoxins

Antibodies cross-reactive with 4 major aflatoxins were demonstrated three weeks after immunization of rabbits with an immunogen which was prepared by conjugating aflatoxin B₃ to bovine serum albumin. Aflatoxin B₃ was first converted to its hemisuccinate before conjugation to the protein. Tritiated aflatoxin B₁ (AFB₁) was used as the marker ligand both for antibody titer determination as well as for analysis of antibody specificity. Competitive RIA revealed that the antibodies have good cross-reactivity with aflatoxins B₁, B₂, G₁, and G₂ when tritiated AFB₁ was used as the marker ligand. The concentrations causing 50% inhibition of binding of³H-AFB₁ to the antibodies by unlabeled aflatoxins B₁, B₂, G₁, G₂ and B₃ were found to be 0.25, 3.34, 0.32, 4.0 and 0.53 ng/assay, respectively. The antibodies could be used for simultaneous analysis of aflatoxins B₁ and G₁, two of the most important toxic metabolites produced byAspergillus flavus andA. parasiticus.     

Production of antibodies against bradykinin

Summary High-titer antibodies against bradykinin were raised in rabbits. 2 different conjugates of bradykinin were used for immunization: bradykinin coupled to human serum albumin via 1,5-difluoro-2,4-dinitrobenzene and bradykinin coupled to edestin via 1-ethyl-3(3-dimethyl-aminopropyl)-carbodiimide. The sensitivity of the radioimmunoassay method is in the range of 1–50 pg of bradykinin. Cross-reaction of anti-bradykinin antisera occurred with kallidin and met-lysbradykinin.     

Production of antibodies against bradykinin.

High-titer antibodies against bradykinin were raised in rabbits. 2 different conjugates of bradykinin were used for immunization: bradykinin coupled to human serum albumin via 1,5-difluoro-2,4-dinitrobenzene and bradykinin coupled to edestin via 1-ethyl-3(3-dimethyl-aminopropyl)-carbodiimide. The sensitivity of the radioimmunoassay method is in the range of 1-50 pg of bradykinin. Cross-reaction of anti-bradykinin antisera occurred with kallidin and met-lys-bradykinin.     

9,10-Dihydroergotamine: Production of antibodies and radioimmunoassay

Summary Antibodies against 9,10-dihydroergotamine (DHE) were produced by immunizing rabbits with a conjugate of 6-nor-6-carboxymethyl-9,10-dihydroergotamine and bovine serum albumin. A highly specific and sensitive radioimmunoassay for DHE has been developed.We are greatly indebted to Dr.T. Fehr for the synthesis of 6-nor-6-carboxymethyl-9,10-dihydroergotamine, and to Dr.E. Schreier for providing us with 9,10-dihydroergotamine-[13-³H]-base of high specific activity.     

Production and characterization of antibody against aflatoxin M1

Summary Antibody against aflatoxin M₁ was obtained after immunization of rabbits with bovine serum albumin-afla M₁ oxime conjugate. The antibody has greatest binding efficiency for afla M₁, and was less efficient for afla B₁. Cross-reaction of antibody with aflatoxin Q₁, aflatoxicol, and aflatoxin B_2a was weak. Aflatoxin B₂, G₁, and G₂ and afla B₁-guanine adduct showed almost no cross-reaction with the antibody. The sensitivity of the binding assay for aflatoxin M₁ detection is in the range of 1–10 ng per assay. Detailed methods for the preparation of the conjugate, production of immune serum, and methods for antibody determination are described.Supported by the College of Agricultural and Life Sciences, North Central Regional project NC-129, the University of Wisconsin-Madison, and by Public Health Service research grant number CA 15064 from the National Cancer Institute, NIH.The authors wish to thank Dr R.C. Garner for providing aflatoxin-B-guanine adduct, and Dr Dennis H. Hseih for providing aflatoxicol and aflatoxin Q₁.     

Production and characterization of antibody against aflatoxin M1

Antibody against aflatoxin M1 was obtained after immunization of rabbits with bovine serum albumin-afla M1 oxime conjugate. The antibody has greatest binding efficiency for afla M1, and was less efficient for afla B1. Cross-reaction of antibody with aflatoxin Q1, aflatoxicol, and aflatoxin B2a was weak. Aflatoxin B2, G1, and G2 and afla B1-guanine adducts showed almost no cross-reaction with the antibody. The sensitivity of the binding assay for aflatoxin M1 detection is in the range of 1-10 ng per assay. Detailed methods for the preparation of the conjugate, production of immune serum, and methods for antibody determination are described.     

Production of porcine antibodies with high specificity to [8-D-arginine] deamino-vasopressin (dDAVP)

Summary The development of a specific radioimmunoassay for [8-D-arginine] deamino-vasopressin (dDAVP) is described.Acknowledgment. We wish to express our thanks to Drs I. Bláha, M. Zaoral, M. Flegel and K. Jot for generous gift of hormones and their analogues used in this study.     

Metabolism of aflatoxins B1 and G1 byAspergillus parasiticus

Résumé Un examen préliminaire fait avec une préparation d'Aspergillus parasiticus est rendu évidente la conversion de l'aflatoxine B en G. Il est probable que l'aflatoxines B₁ est le précurseur biogénétique d'autres aflatoxines produite par le champignon.

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K. K. M. is grateful to the Council of Scientific and Industrial Research, New Delhi, India, for the award of a Senior Research Fellowship.     

Production of antibodies to amanitins as the basis for their radioimmunoassay

Summary The production of antibodies against amanitins is described. By means of these antibodies, a radioimmunoassay was developed which allows detection of as little as 0.5 ng of amanitins in 1 ml of serum. By this method, the clearance of α-amanitin from the blood of poisoned mice was measured. Acknowledgments. We thank ProfessorT. Wieland and Dr.H. Faulstich (Heidelberg) for their generous gift of β-amanitin and [³H]O-methyl-demethyl-γ-amanitin. This work was supported by grants from C.N.R., Rome.     

Production of antibodies against synthetic angiotensin II in various animal species

Zusammenfassung Bei Ratten und Kaninchen, jedoch nicht bei Hunden und Katzen, konnten durch wiederholte Injektion von Angiotensin-II-Amid-Carbodiimid-Proteinkomplex in Freund'schem Adjuvans zirkulierende Antikörper gegen Angiotensin erzeugt werden. Durch diese Immunisierung wurde die pressorische Wirkung von exogenem Angiotensin aufgehoben bzw. abgeschwächt, während die von Noradrenalin oder Vasopressin unbeeinflusst blieb. Der Reningehalt der Nieren von Ratten und Kaninchen wurde durch die Immunisierung nicht verändert.     

Biosynthesis of aflatoxins by cell-free preparations fromAspergillus flavus

Résumé Incorporation d'acétate-1-¹⁴C, de leucine-U-¹⁴C et de mévalonate-2-¹⁴C sous forme d'aflatoxine dans les extraits libres des cellulesd'Aspergillus flavus. Dans le cas du mévalonate, l'incorporation de l'ordre de 0,4 pour 100 et les activités spécifiques sont 13 000 à 40 000 cpm/mg d'aflatoxine. La capacité d'incorporation des substrats en aflatoxine s'observe, en particulier, dans les mitochondries.     

Production of antibodies to amanitins as the basis for their radioimmunoassay.

The production of antibodies against amanitins is described. By means of these antibodies, a radioimmunoassay was developed which allows detection of as little as 0.5 ng of amanitins in 1 ml of serum. By this method, the clearance of alpha-amanitin from the blood of poisoned mice was measured.     

Binding of aflatoxins B1 and G1 to human serum proteins

Zusammenfassung In vitro Experimente mit¹⁴C-markiertem, gereinigtem Aflatoxin zur Untersuchung der Bindung von Aflatoxin B₁ und G₁ an verschiedene Serumproteine ergaben, dass Aflatoxin B₁ hauptsächlich mit-Globulin, G₁ dagegen vorwiegend mit Albumin bindet.

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This work was supported by part by a grant from the China Medical Board of New York, Inc., and was performed during one of us (S.S.L.) received a class C. research award from the National Science Council, Republic of China.     

Production of monoclonal anti-Schistosoma mansoni antibodies. Preliminary study of their biological activities]

Monoclonal antibodies against Schistosoma mansoni have been produced by fusion of splenic lymphocytes from S. mansoni infected Rats and P3-X63-Ag8 BALB/c cells. In vitro and in vivo studies of the biological activities of these antibodies have led to the identification of IgE antibodies with a high reaginic activity and antibodies which in a complement dependent or eosinophil dependent system were shown to have a marked cytotoxicity for schistosomula in vitro. This methodology seems to open new perspectives for the study of antibody function in immunity against parasites as well as for the isolation of the corresponding target antigens.