Abstract
In order to clarify the specificity of serum sialyltransferase in patients with rheumatoid arthritis (RA), we studies the serum which was free of platelet components, by isoelectric focusing. Isoelectric focusing was performed using isoelectric focusing apparatus (110 ml capacity) . 2.5% ampholine (pH 3-10) were used in a gradient of 0-50 w/v% sucrose. The starting amperage was 3.0mA and 800V. After 23 hours the voltage was adjusted to 1400V. Electrofocusing was conducted for 47 hours at 0-1°C. After electrofocusing 2.0-ml fractions were collected. 30-μl aliquots of each fraction were assayed for asialofetuin sialyltransferase activity using CMP- (9-3H) sialic acid as substrate. Isozymic forms were showed at pI 4.20, 4.65, 5.10, 6.20 and 6.80 in serum from patient with RA. Platelet lysate showed a large peak of ST activity at pI. 6.20.Concludingly, we found by isoelectric focusing that acid ST isozymic forms were in serum and that platelet-derived ST (pI 6.20) was contaminated in circulating plasma.
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