Characterization and measurement of human apolipoprotein A-II by radioimmunoassay
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 det...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
1980-09-01
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| Series: | Journal of Lipid Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S002222752034788X |
| _version_ | 1818671489447100416 |
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| author | R B Goldberg J B Karlin D J Juhn A M Scanu C Edelstein A H Rubenstein |
| author_facet | R B Goldberg J B Karlin D J Juhn A M Scanu C Edelstein A H Rubenstein |
| author_sort | R B Goldberg |
| collection | DOAJ |
| description | 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 non-iodinated 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. |
| first_indexed | 2024-12-17T07:24:49Z |
| format | Article |
| id | doaj.art-8b0c2ae3b4fa40018755c2db0151cc3a |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-17T07:24:49Z |
| publishDate | 1980-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-8b0c2ae3b4fa40018755c2db0151cc3a2022-12-21T21:58:39ZengElsevierJournal of Lipid Research0022-22751980-09-01217902912Characterization and measurement of human apolipoprotein A-II by radioimmunoassayR B GoldbergJ B KarlinD J JuhnA M ScanuC EdelsteinA H RubensteinThe 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 non-iodinated 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.http://www.sciencedirect.com/science/article/pii/S002222752034788X |
| spellingShingle | R B Goldberg J B Karlin D J Juhn A M Scanu C Edelstein A H Rubenstein Characterization and measurement of human apolipoprotein A-II by radioimmunoassay Journal of Lipid Research |
| title | Characterization and measurement of human apolipoprotein A-II by radioimmunoassay |
| title_full | Characterization and measurement of human apolipoprotein A-II by radioimmunoassay |
| title_fullStr | Characterization and measurement of human apolipoprotein A-II by radioimmunoassay |
| title_full_unstemmed | Characterization and measurement of human apolipoprotein A-II by radioimmunoassay |
| title_short | Characterization and measurement of human apolipoprotein A-II by radioimmunoassay |
| title_sort | characterization and measurement of human apolipoprotein a ii by radioimmunoassay |
| url | http://www.sciencedirect.com/science/article/pii/S002222752034788X |
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