Immunoturbidimetric determination of lipoprotein(a): improvement in the measurement of turbid and triglyceride-rich samples.
Immunoturbidimetry (IT), a widely used method in clinical chemical laboratories, was checked for its suitability for lipoprotein(a) (Lp(a)) quantification. When the conventional sample diluents were used, turbidimetry gave false results particularly with frozen or lipemic sera which correlated poorly with electroimmunodiffusion (EID). L-Proline which is known to dissociate Lp(a) from other apo B-containing lipoproteins improved the results considerably. One hundred frozen sera were investigated in IT with and without the addition of L-proline to the sample diluent. EID served as a comparison method. In a method comparison (IT vs. EID) linear regression analysis improved from r = 0.793: y = 0.89x - 9.4 (without L-proline) to r = 0.949: y = 0.98x + 4.8 (with L-proline). The improvement of the correlation of the two methods was most pronounced in sera with triglyceride values exceeding 5.5 mmol/l. The IT assay described here was linear between 50 and 1100 mg/l. Total imprecision (coefficient of variation) was below 10%. The assay was not affected by the addition of LDL or plasminogen to the samples. The Lp(a) concentration of the calibrator, i.e. a secondary standard serum, was compared with that of a purified primary Lp(a) standard which consisted of a mixture of four apo(a) isoforms. Total Lp(a) mass (lipids, protein, carbohydrates) was determined chemically and was compared with the Lp(a) mass determined immunochemically by IT and EID. Recovery of the purified Lp(a) was 106% (range 90-116%) in IT and 102% (range 91-115%) in EID. Dose response curves from pure single isoforms (S1 and S4), calibrator and serum samples were parallel. We consider IT to be a simple and rapid method for Lp(a) quantification which is not biased by different apo(a) isoforms.
Molinari
E A
EA
IMMUNO AG, Vienna, Austria.
Pichler
P F
PF
Grillhofer
H
H
Kostner
G M
GM
D001798:Blood Proteins / Q000032:analysis; D002241:Carbohydrates / Q000097:blood; D002784:Cholesterol / Q000097:blood; D006801:Humans; D005779:Immunodiffusion / Q000379:methods; D017270:Lipoprotein(a) / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D008077:Lipoproteins, LDL / Q000097:blood; D009391:Nephelometry and Turbidimetry / Q000379:methods*; D010743:Phospholipids / Q000097:blood; D010958:Plasminogen / Q000032:analysis; D011392:Proline / Q000494:pharmacology; D012015:Reference Standards; D015203:Reproducibility of Results; D014280:Triglycerides / Q000097:blood
D001798:Blood Proteins; D002241:Carbohydrates; D017270:Lipoprotein(a); D008077:Lipoproteins, LDL; D010743:Phospholipids; D014280:Triglycerides; D010958:Plasminogen; D002784:Cholesterol; D011392:Proline