Immunoaffinity purification of the lipid transfer protein complex directly from human plasma
| Autor: | Steven J. Busch, Jeff L. Ellsworth, Jahei Ihm, Charles R. Duvic, Judith A.K. Harmony |
|---|---|
| Rok vydání: | 1986 |
| Předmět: |
Adult
food.ingredient Sterol O-acyltransferase Biophysics Biochemistry Lecithin Chromatography Affinity Sepharose chemistry.chemical_compound food Humans Molecular Biology Chromatography biology Immunochemistry Albumin Cell Biology Molecular Weight Isoelectric point chemistry biology.protein Cholesteryl ester Antibody Carrier Proteins Plant lipid transfer proteins |
| Zdroj: | Analytical biochemistry. 153(1) |
| ISSN: | 0003-2697 |
| Popis: | The human cholesteryl ester (CE) and triglyceride (TG) exchange protein (denoted LTC or lipid transfer complex) was isolated in a single step from plasma using immunoaffinity batch extraction. Antibodies were raised against two preparations of conventionally purified LTC. LTC-I and LTC-II (purified 20,000-fold and 3500-fold, respectively) were used as immunogens. The antiLTC antibodies were isolated by anion-exchange chromatography and coupled to Affi-Gel 10. Chromatography of plasma on antiLTC Affi-Gel removed all of the CE and TG transfer activity. Moreover, LTC prepared from both antiLTC-I and antiLTC-II-Affi-Gel matrices were identical when analyzed by sodium dodecyl sulfate-polyacrylamide gel LTC electrophoresis. LTC exhibited two protein bands of Mr (apparent) 67,000 and 58,000 and a broad, faintly staining region at greater than 150,000. Analysis of LTC by immunoblotting indicated that both antiLTC-I and antiLTC-II antibodies recognized the same LTC proteins. Isoelectric focussing of LTC gave two pI values, 5.2 and 8.7. These data suggest that LTC is a complex of specific proteins and perhaps lipid. Specific CE and TG exchange activities of immunoaffinity-purified LTC were comparable, although the activities were low with respect to that of the antigen used to generate antiLTC-I. This is not due to contamination of LTC by albumin, lecithin:cholesterol acyltransferase, or apolipoproteins AI, AII, B, CIII, D, or E. |
| Databáze: | OpenAIRE |
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