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Study of human cerebrospinal fluid proteins by size exclusion‐high performance liquid chromatography and two‐dimensional gel electrophoresis
Author(s) -
Wiederkehr Felix,
Büeler Martin R.,
Wacker Marianne,
Vonderschmitt Dieter J.
Publication year - 1989
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150100706
Subject(s) - chemistry , chromatography , size exclusion chromatography , high performance liquid chromatography , polyclonal antibodies , fraction (chemistry) , gel electrophoresis , transthyretin , monomer , cerebrospinal fluid , biochemistry , antibody , biology , polymer , organic chemistry , neuroscience , immunology , enzyme , endocrinology
Cerebrospinal fluid (CSF) proteins were separated into three main fractions by size exclusion‐high performance liquid chromatography (SE‐HPLC). Subsequent analysis of each fraction by two‐dimensional gel electrophoresis (2‐DE) facilitated the detection of trace components in CSF and additionally provided more information about the native properties of various proteins. Certain proteins are present in a polymeric form and appear in the high molecular weight SE‐HPLC fraction. In the middle molecular weight SE‐HPLC fraction we found a CSF‐specific transthyretinrelated protein by immunoblotting with polyclonal antibodies to transthyretin. Possible interpolypeptide disulfide bonds of such polymeric proteins were studied using a nonreducing 2‐DE system. This procedure revealed that all apolipoprotein E monomers in CSF, which are synthesized in astrocytes, are linked by disulfide bonds. In the CSF from a patient with clinically definite multiple sclerosis (MS), novel proteins appeared in the high molecular weight SE‐HPLC fraction, which are obscured by other proteins if total CSF is analyzed.