Abstract
A radioimmunoassay (RIA) for vitamin B12 is described. Antisera were raised in rabbits using a conjugate between 5′-O-succinyl cyanocobalamin and chicken serum albumin. The antibody is largely specific for the β (upper) face of the vitamin B12 molecule. The RIA can detect vitamin B12 down to concentrations of 37 pmol/L and correlated well (r=0.980, p<0.001) with a commercial radioassay. The use of this RIA, together with an assay for plasma methylmalonic acid, in the diagnosis of cobalt/vitamin B12 deficiency in sheep is described.
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Ahrenstedt, S.S. and Thorell, J.I., 1979. The production of antibodies to vitamin B12. Clinica Chimica Acta, 95, 419–423
Allen, R.H., 1981. Current status of cobalamin (vitamin B12) assays. Syllabus of the 7th Annual Meeting of the Clinical Radioassay Society, pp. 85–109
Allen, R.H. and Majerus, P.W., 1972. Isolation of vitamin B12 binding proteins using affinity chromatography. Journal of Biological Chemistry, 247, 7695–7701
Endres, D.B., Painter, K. and Niswender, G.D., 1978. A solid phase radioimmunoassay for vitamin B12 in serum, with use of radioiodinated tyrosine methyl ester of vitamin B12. Clinical Chemistry, 24, 460–465
Gershman, H., Nathanson, N., Abeles, R.H. and Levine, L., 1972. Production and specificity of antibodies to B12 derivatives. Archives of Biochemistry and Biophysics, 153, 407–409
Green, R., Newmark, P.A., Musso, A.M. and Mollin, D.L., 1974. The use of chicken serum for measurement of serum vitamin B12 concentration by radioisotope dilution. British Journal of Haematology, 27, 507–526
Hurn, B.A.L. and Chantler, S.M., 1980. Production of reagent antibodies. Methods in Enzymology, 70, 104–142
Lau, K.S., Gottlieb, C., Wasserman, L.R. and Herbert, V., 1965. Measurement of serum vitamin B12 using radioisotope dilution and coated charcoal. Blood, 26, 202–214
Lindenbaum, J., Healton, E.B., Savage, D.G., Brust, J.C.M., Garrett, T.J., Podell, E. R., Marcell, P.D., Stabler, S.P. and Allen, R.H., 1988. Neuropsychiatric disorders caused by cobalamin deficiency in the absence of anemia or macrocytosis. New England Journal of Medicine, 318, 1720–1728
Linnell, J.C., Collings, L., Down, M.C. and England, J.M., 1979. Distribution of endogenous cobalamin between the transcobalamins in various mammals. Clinical Science, 57, 139–144
Lowry, O.H., Rosenbrough, N.J., Farr, A.L. and Randall, R.J., 1951. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193, 265–275
McMurray, C.H., Rice, D.A., McLoughlin, M. and Blanchflower, W.J., 1985. Cobalt deficiency and the potential of using methylmalonic acid as a diagnostic and prognostic indicator. In C.F. Mills, I. Bremner and J.K. Chesters (eds.), Trace Elements in Man and Animals, Vol. 5, (CAB, UK) pp. 603–608
McMurray, C.H., Blanchflower, W.J., Rice, D.A. and McLoughlin, M., 1986. Sensitive and specific gas chromatographic method for the determination of methylmalonic acid in plasma and urine of ruminants. Journal of Chromatography, 378, 201–207
Mollin, D.L., Hoffbrand, A.V., Ward, P.G. and Lewis, S.M., 1980. Interlaboratory comparison of serum vitamin B12 assay. Journal of Clinical Pathology, 33, 243–248
Nexo, E. and Olesen, H., 1982. Quantitation of cobalamins in human serum. In D. Dolphin (ed.), B 12, (Wiley Interscience, New York) pp. 87–104
O'Harte, F.P.M., Kennedy, D.G., Blanchflower, W.J. and Rice, D.A., 1989. Methylmalonic acid in the diagnosis of cobalt deficiency in barley fed lambs. British Journal of Nutrition, 62, 729–738
Pratt, J.J. and Woldring, M.G., 1982. Radioassay of vitamin B12 and other corrinoids. Methods in Enzymology, 84, 369–406
Quadros, E.V., Rothenberg, S.P. and Polu, S., 1988. A specific radioimmunoassay for 5′-deoxyadenosyl cobalamin in serum. British Journal of Haematology, 69, 551–557
Rice, D.A., McLoughlin, M., Blanchflower, W.J., Goodall, E.A. and McMurray, C.H., 1987. Methylmalonic acid as an indicator of vitamin B12 deficiency in grazing sheep. Veterinary Record, 121, 472–473
Rosenthal, H.L. and Sarett, H.P., 1952. The determination of vitamin B12 activity in human serum. Journal of Biological Chemistry, 199, 433–442
Ross, G.I.M., 1950. Vitamin B12 assay in body fluids. Nature, 166, 270–271
Rothenberg, S.P., Marcoullis, G.P., Schwaz, S. and Lader, E., 1984. Measurement of cyanocobalamin in serum by a specific radioimmunoassay. Journal of Laboratory and Clinical Medicine, 103, 959–972
Scatchard, G., 1949. The attractions of proteins for small molecules and ions. Annals of the New York Academy of Science, 51, 660–672
Toraya, T., Ohashi, K., Ueno, H. and Fukui, S., 1974. Succinyl derivatives of vitamin B12. Vitamins (Japan), 48, 557–563
Toraya, T., Ohashi, K., Ueno, H. and Fukui, S., 1975. Preparation, properties and biological activities of succinyl derivatives of vitamin B12. Bioinorganic Chemistry, 4, 245–255
Van de Wiel, D.F.M., Goedemans, W.T. and Woldring, M.G., 1974. Production and purification of antibody against protein-vitamin B12 conjugates for radioimmunoassay purposes. Clinica Chimica Acta, 56, 143–149
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Kennedy, D.G., O'Harte, F.P.M., Blanchflower, W.J. et al. Development of a specific radioimmunoassay for vitamin B12 and its application in the diagnosis of cobalt deficiency in sheep. Veterinary Research Communications 14, 255–265 (1990). https://doi.org/10.1007/BF00350708
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DOI: https://doi.org/10.1007/BF00350708