Determination of human apolipoprotein C-II by electroimmunoassay. Studies on standardization and determination before and after physical training

• Published in: International journal of biochemistry Vol. 15; no. 4; p. 501
• Main Authors: Jauhiainen, M, Laitinen, M, Penttilä, I, Puhakainen, E, Hietanen, E
• Format: Journal Article
• Language: English
• Published: England 1983
• Subjects: Adult; Apolipoprotein C-II; Apolipoproteins - blood; Apolipoproteins - isolation & purification; Apolipoproteins C; Female; Humans; Immunodiffusion; Immunoelectrophoresis; Lipoproteins, HDL - blood; Lipoproteins, VLDL - blood; Male; Middle Aged; Physical Exertion
• ISSN: 0020-711X
• DOI: 10.1016/0020-711X(83)90123-4

1. Human VLDL and HDL were fractionated by sequential ultracentrifugation until free of contaminant plasma proteins. 2. Column chromatofocusing method was used to isolate apolipoprotein C-II from apoVLDL and apo HDL. C-apoprotein peak was rechromatofocused and the second peak was the apo C-II (pI 4.7, homogeneous band on SDS slab gel). 3. New Zealand white rabbits were immunized with apo C-II. Antiserum gave a single precipitate are of identity between whole serum, apoVLDL, apoHDL and apo C-II. 4. Apo C-II concentration was measured by electroimmunoassay method. During standardization 1% Triton X-100 improved the rocket shapes and contours. Total delipidation did not affect the assay system and so the antigenic determinants of apo C-II are all available to antiserum. The lowest concentration of apo C-II possible to determine with this method was 70 ng/sample well. 5. There was no difference between the apo C-II values before (39.8 +/- 7.1 mg/l, n = 19) and after (41.6 +/- 6.4 mg/l, n = 19) moderate physical training among normolipemic subjects. 6. Specific immunoprecipitation technique was also used to determine apo C-II content in standard pool serum.