Characterization of antibodies to chicken riboflavin carrier protein: Antigenicity of the tryptic fragments |
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Authors: | Usha Natraj K S N Iyer Vijaya Raghavan Smita Mahale Jacintha Pereira |
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Institution: | (1) Institute for Research in Reproduction, Jehangir Merwanji Street, 400 012 Parel, Bombay, India |
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Abstract: | The antigenecity of tryptic fragments of reduced and carboxymethylated chicken riboflavin carrier protein were studied. The
tryptic sites of the native riboflavin carrier protein bound to riboflavin were inaccessible. The molecular weight and the
elution profile on high performance liquid chromatography (TSK 545 DEAE) were unaltered at an enzyme to substrate ratio of
1:31. However, carboxymethylated riboflavin carrier protein could be cleaved into 3 or 4 fragments at an enzyme to substrate
ratio of 1:250 or 1:125. Chromatographic separation of the tryptic fragments on high pressure liquid chromatography (TSK 545
DEAE) revealed the presence of two fragments with different elution profiles but similar molecular weight 26 ±2 kDa. Only
one fragment (associated with peak 2) had the ability to displace chicken riboflavin carrier protein in an homologous chicken
riboflavin carrier protein radioimmunoassay. Thus, carboxymethylated ribotlavin carrier protein which does not compete with
chicken riboflavin carrier protein in the radioimmunoassay, on mild trypsinization generates a fragment which interacts with
chicken riboflavin carrier protein in radioimmunoassay. |
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Keywords: | Riboflavin carrier protein tryptic fragments antigenicity |
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