Characterization and measurement of human apolipoprotein A-II by radioimmunoassay

Ronald B Goldberg, J. B. Karlin, D. J. Juhn, A. M. Scanu, C. Edelstein, A. H. Rubenstein

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The development of a radioimmunoassay for apolipoprotein A-II (apo A-II) is described. Initial studies revealed a lack of immunological identity between purified apo A-II used as the standard and serum or HDL. Extensive testing of different buffers, standards, antisera, tracers, utilization of a detergent, and heating of sera failed to resolve the problem. Gel filtration of iodinated and noniodinated apo A-II on Sephadex G-100 columns showed that apo A-II, in dilute solution, elutes in a higher molecular zone than expected with a broad, assymetrical profile. The use of a subfraction of the tracer in the assay resulted in parallelism in the serum and standard dilution curves. The apo A-II assay was sensitive, specific and reproducible. Apo A-II added to sera was fully recovered and delipidation did not affect the immunoreactivity of either serum or HDL. Apo A-II contributed approximately 20% to the protein mass of HDL. Comparison of these results with those obtained by radial immunodiffusion, and with previously reported data, indicates that the reactivity of apo A-II in its native and delipidated forms may be markedly influenced by different immunologic methodologies and their specific reagents. Caution should thus be shown at present in assigning absolute concentrations to apo A-II in serum or HDL.

Original languageEnglish
Pages (from-to)902-912
Number of pages11
JournalJournal of Lipid Research
Volume21
Issue number7
StatePublished - Dec 1 1980
Externally publishedYes

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Apolipoprotein A-II
Radioimmunoassay
Serum
Assays
human APOA2 protein
Immunodiffusion
Detergents
Heating
Dilution
Gel Chromatography
Immune Sera
Buffers
Gels

ASJC Scopus subject areas

  • Endocrinology

Cite this

Goldberg, R. B., Karlin, J. B., Juhn, D. J., Scanu, A. M., Edelstein, C., & Rubenstein, A. H. (1980). Characterization and measurement of human apolipoprotein A-II by radioimmunoassay. Journal of Lipid Research, 21(7), 902-912.

Characterization and measurement of human apolipoprotein A-II by radioimmunoassay. / Goldberg, Ronald B; Karlin, J. B.; Juhn, D. J.; Scanu, A. M.; Edelstein, C.; Rubenstein, A. H.

In: Journal of Lipid Research, Vol. 21, No. 7, 01.12.1980, p. 902-912.

Research output: Contribution to journalArticle

Goldberg, RB, Karlin, JB, Juhn, DJ, Scanu, AM, Edelstein, C & Rubenstein, AH 1980, 'Characterization and measurement of human apolipoprotein A-II by radioimmunoassay', Journal of Lipid Research, vol. 21, no. 7, pp. 902-912.
Goldberg RB, Karlin JB, Juhn DJ, Scanu AM, Edelstein C, Rubenstein AH. Characterization and measurement of human apolipoprotein A-II by radioimmunoassay. Journal of Lipid Research. 1980 Dec 1;21(7):902-912.
Goldberg, Ronald B ; Karlin, J. B. ; Juhn, D. J. ; Scanu, A. M. ; Edelstein, C. ; Rubenstein, A. H. / Characterization and measurement of human apolipoprotein A-II by radioimmunoassay. In: Journal of Lipid Research. 1980 ; Vol. 21, No. 7. pp. 902-912.
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