Preparativeseparationof immunoglobulins from bovine colostrum by continuous divergent-flow electrophoresis

0562904 - ÚIACH 2024 RIV DE eng J - Journal Article
Šťastná, Miroslava - Šlais, Karel
Preparativeseparationof immunoglobulins from bovine colostrum by continuous divergent-flow electrophoresis.
Journal of Separation Science. Roč. 46, č. 1 (2023), č. článku 2200679. ISSN 1615-9306. E-ISSN 1615-9314
Institutional support: RVO:68081715
Keywords : continuous flow preparation * isotachophoresis * moving boundary electrophoresis
OECD category: Analytical chemistry
Impact factor: 3.1, year: 2022
Method of publishing: Limited access

Immunoglobulins in bovine colostrum were separated and fractionated from other proteins using method and instrumentation developed in our laboratory. The proposed separation was based on bidirectional isotachophoresis/moving boundary electrophoresis with electrofocusing of the analytes in pH gradient from 3.9 to 10.1. The preparative instrumentation included the trapezoidal non-woven fabric that served as separation space with divergent continuous flow. The defatted and casein precipitate free colostrum supernatant was loaded directly into instrument without any additional colostrum pre-preparation. Immunoglobulin G that was fractionated from other immune proteins such as bovine serum albumin, β-lactoglobulin and α-lactalbumin, was continuously collected in separated fractions over 3 hours. The fractions were further processed, and isolated immunoglobulin G in the liquid fractions was confirmed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and by re-focusing in gel isoelectric focusing. Separated immunoglobulin G was detected in seven fractions by sodium dodecyl sulphate-polyacrylamide gel electrophoresis with gradually decreased concentration in the fractions. Re-focusing of the proteins in the fractions by gel isoelectric focusing revealed multiple separated zones of immunoglobulin G with the pI values covering the range from 5.4 to 7.2. Each fraction contained the distinct zones with gradually increased pI values and decreased concentrations from fraction to fraction.
Permanent Link: https://hdl.handle.net/11104/0335073