Immunoassay utilizing .sup.125 I Protein A

Drug – bio-affecting and body treating compositions – Radionuclide or intended radionuclide containing; adjuvant...

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260112R, 435883, 436501, 436518, 436520, 436539, 436542, 436828, 436804, G01N 3348, G01N 3360

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044303181

ABSTRACT:
An improved method for the preparation of .sup.125 I-labelled Protein A (.sup.125 I PA) of high specific and functional activity. .sup.125 I PA has been used in combination with purified rabbit IgG (immunoglobulin G) bound to a solid support to develop a competitive binding assay capable of detecting Protein A or human, rabbit and guinea pig IgG at the nanogram level. Additionally, .sup.125 I PA may be used to detect methotrexate, leucovorin and similar substances.
.sup.125 I PA has also been used to detect IgG anti-Forssman antibody bound to sheep erythrocytes and to line-1 and line-10 tumor cells and as an indirect assay for tumor associated antigen in the ascitic fluid of tumor-bearing guinea pigs.
Additionally, an improved method of preparation of iodination of Protein A is utilized. This procedure used the Bolton-Hunter (1973) reagent of radioactive iodine in benzene which carrier is evaporated. The PA is added or attached while in an amino acid mixture and separation is per column utilizing Sephadex G-25 and VBS-gel.

REFERENCES:
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patent: 4230685 (1980-10-01), Senyei et al.
Langone, John J., Methods in Enzymology, vol. 70, pp. 356-375, Academic Press (1980).
Bolton, A. E. and Hunter, W. M., Biochemical Journal, vol. 133, p. 529 (1973).
Langone, John J. et al., Journal of Immunological Methods, vol. 18, pp. 281-293 (1978), vol. 24, pp. 269-285 (1978).
Dorval, G. et al., J. Immunological Methods, vol. 7, p. 237 (1975).
Meltzer, P. M. et al., Journal of Immunological Methods, vol. 17, p. 163 (1977).
Bikerfeld, P. et al., Journal of Immunological Methods, vol. 6, p. 249 (1975).
Hunter, W. M. et al., Nature (London) vol. 194, p. 495 (1962).

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