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March 1, 1976Clinical Chemistry288 citationsOpen Access

Determination of apolipoprotein A and its constitutive A-I and A-II polypeptides by separate electroimmunoassays.

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MCMichael D. CurryPAPetar AlaupovicCSC. A. Suenram

Key Points

  • To develop and validate electroimmunoassays for quantifying apolipoprotein A and its associated polypeptides A-I and A-II.
  • Developed electroimmunoassays for measuring apolipoprotein A, A-I, and A-II in human serum.
  • Used purified lipoprotein A, A-I, and A-II to prepare monospecific antisera.
  • Assays demonstrated high precision and accuracy with rapid results in serum and various lipoprotein density classes.
  • Normal serum concentrations for A-I were 143 +/- 24 mg/dl and A-II were 78 +/- 17 mg/dl.
  • Patients with type I disease had significantly lower concentrations of A-I (0.3-30 mg/dl) and A-II (11-20 mg/dl).
  • The molar ratio of A-I to A-II in serum and high-density lipoproteins was close to 1.0.

Abstract

Electroimmunoassays ("rocket" electrophoresis) are described for human serum apolipoprotein A and its constitutive A-I and A-II polypeptides. Purified lipoprotein A, A-I, and A-II were used to prepare monospecific antisera and to standardize assays. These specific, rapid (5-8 h), precise (the within-and between-assay coefficients of variations are 5 and 7%, respectively), and accurate (by gravimetry) assays are applicable to measurement of these polypeptides in whole serum and in various density classes of lipoproteins. Comparable results are obtained with intact and delipidized lipoproteins. Results correlated well with those obtained by radial immunodiffusion or radioimmunoassay. However, the present procedure is more rapid than the former and simpler than the latter immunoassay. Concentrations of A-I and A-II in the serum of normal men and women were similar (143 +/- 24 and 146 +/- 78 mg/dl, respectively, for A-I and 78 +/- 17 and 83 +/- 25 mg/dl for A-II). Subjects with type lla, llb, and IV hyperlipoproteinemias had similar concentrations of both polypeptides, while patients with type I disease, lecithin:cholesterol acyltransferase deficiency and LP-A deficiency had lowest concentrations of A-I (0.3-30 mg/dl) and A-II (11-20 mg/dl). The molar ratio of A-I/A-II in the serum and high-density lipoproteins was close to unity.

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Cite This Study

Curry et al. (1976) studied this question.

synapsesocial.com/papers/6a6b5e7a6f523709a0249a8dhttps://doi.org/10.1093/clinchem/22.3.315
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