A comparison of the properties of chicken serum with other vitamin B12 binding proteins used in radioisotope dilution methods for measuring serum vitamin B12 concentrations

P. A. Newmark, Ralph Green, A. M. Musso, D. L. Mollin

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The effects of pH, ionic strength, and vitamin B12 concentration on the vitamin B12 binding capacities of chicken serum, normal human serum, chronic myelogenous leukaemia serum, and hog intrinsic factor concentrates are compared. The influence of pH on B12 binding capacity was shown to be greatest at low pH and that of ionic strength was most evident below 0.1 ionic strength, particularly at acidic pH. The effect of B12 concentration of B12 binding capacity was shown to vary with different binders, the pH of binding, and the time of incubation. It was possible to select conditions for carrying out the binding of B12 to chicken serum so that the amount of B12 bound was virtually unaffected by the kind of variations in pH, ionic strength, or concentration of B12 that could occur in a radioisotope dilution assay of serum vitamin B12. Under these conditions it was further demonstrated that the binding was unaffected by the presence of denatured human serum. In addition, chicken serum had a very great binding capacity and affinity for B12, was very stable on storage and was compatible with the use of protein coated charcoal. Some conclusions on the optimal use of B12 binding protein in the radioisotope dilution assay of serum vitamin B12 are made and the high potentiality of chicken serum in such a system is suggested.

Original languageEnglish (US)
Pages (from-to)359-373
Number of pages15
JournalBritish Journal of Haematology
Volume25
Issue number3
StatePublished - 1973
Externally publishedYes

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Transcobalamins
Vitamin B 12
Radioisotopes
Chickens
Serum
Osmolar Concentration
Intrinsic Factor
Charcoal
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Carrier Proteins

ASJC Scopus subject areas

  • Hematology

Cite this

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title = "A comparison of the properties of chicken serum with other vitamin B12 binding proteins used in radioisotope dilution methods for measuring serum vitamin B12 concentrations",
abstract = "The effects of pH, ionic strength, and vitamin B12 concentration on the vitamin B12 binding capacities of chicken serum, normal human serum, chronic myelogenous leukaemia serum, and hog intrinsic factor concentrates are compared. The influence of pH on B12 binding capacity was shown to be greatest at low pH and that of ionic strength was most evident below 0.1 ionic strength, particularly at acidic pH. The effect of B12 concentration of B12 binding capacity was shown to vary with different binders, the pH of binding, and the time of incubation. It was possible to select conditions for carrying out the binding of B12 to chicken serum so that the amount of B12 bound was virtually unaffected by the kind of variations in pH, ionic strength, or concentration of B12 that could occur in a radioisotope dilution assay of serum vitamin B12. Under these conditions it was further demonstrated that the binding was unaffected by the presence of denatured human serum. In addition, chicken serum had a very great binding capacity and affinity for B12, was very stable on storage and was compatible with the use of protein coated charcoal. Some conclusions on the optimal use of B12 binding protein in the radioisotope dilution assay of serum vitamin B12 are made and the high potentiality of chicken serum in such a system is suggested.",
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AU - Newmark, P. A.

AU - Green, Ralph

AU - Musso, A. M.

AU - Mollin, D. L.

PY - 1973

Y1 - 1973

N2 - The effects of pH, ionic strength, and vitamin B12 concentration on the vitamin B12 binding capacities of chicken serum, normal human serum, chronic myelogenous leukaemia serum, and hog intrinsic factor concentrates are compared. The influence of pH on B12 binding capacity was shown to be greatest at low pH and that of ionic strength was most evident below 0.1 ionic strength, particularly at acidic pH. The effect of B12 concentration of B12 binding capacity was shown to vary with different binders, the pH of binding, and the time of incubation. It was possible to select conditions for carrying out the binding of B12 to chicken serum so that the amount of B12 bound was virtually unaffected by the kind of variations in pH, ionic strength, or concentration of B12 that could occur in a radioisotope dilution assay of serum vitamin B12. Under these conditions it was further demonstrated that the binding was unaffected by the presence of denatured human serum. In addition, chicken serum had a very great binding capacity and affinity for B12, was very stable on storage and was compatible with the use of protein coated charcoal. Some conclusions on the optimal use of B12 binding protein in the radioisotope dilution assay of serum vitamin B12 are made and the high potentiality of chicken serum in such a system is suggested.

AB - The effects of pH, ionic strength, and vitamin B12 concentration on the vitamin B12 binding capacities of chicken serum, normal human serum, chronic myelogenous leukaemia serum, and hog intrinsic factor concentrates are compared. The influence of pH on B12 binding capacity was shown to be greatest at low pH and that of ionic strength was most evident below 0.1 ionic strength, particularly at acidic pH. The effect of B12 concentration of B12 binding capacity was shown to vary with different binders, the pH of binding, and the time of incubation. It was possible to select conditions for carrying out the binding of B12 to chicken serum so that the amount of B12 bound was virtually unaffected by the kind of variations in pH, ionic strength, or concentration of B12 that could occur in a radioisotope dilution assay of serum vitamin B12. Under these conditions it was further demonstrated that the binding was unaffected by the presence of denatured human serum. In addition, chicken serum had a very great binding capacity and affinity for B12, was very stable on storage and was compatible with the use of protein coated charcoal. Some conclusions on the optimal use of B12 binding protein in the radioisotope dilution assay of serum vitamin B12 are made and the high potentiality of chicken serum in such a system is suggested.

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