Circumventing Complement C3 Interference in the Analysis of Carbohydrate-Deficient Transferrin in Fresh Serum

✍ Scribed by Amy T. Beisler; Robert H. Kelly; James P. Landers

• Publisher: Elsevier Science
• Year: 2000
• Tongue: English
• Weight: 134 KB
• Volume: 285
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2000.4717

✦ Synopsis

A method for preventing interference by the glycoprotein complement C3 and its ␤-globulin split products in the capillary electrophoretic analysis of carbohydrate-deficient transferrin was developed. Inulin was used to activate the alternate complement pathway and convert native C3 into various degradation products whose electrophoretic mobility no longer coincides with the transferrin glycoforms. Capillary electrophoresis and zone electrophoresis on agarose gel were used to monitor reaction conditions for alternate complement pathway activation. Incubation of 50 L of fresh serum with 180 L of a 50 mg/mL inulin slurry for 12 h removed the native C3 peak from the ␤ region. Inulin treatment did not affect electrophoretic behavior of other ␤-globulins, including transferrin. Altering the electrophoretic behavior of complement C3, by treating fresh serum with inulin, permits rapid capillary electrophoresis evaluation of carbohydratedeficient transferrin glycoforms.