Isolation of Immunoglobulins and Their Use in Immunoaffmity HPLC

Josić, Dj.; Hofmann, Werner; Habermann, R.; Schulzke, JD; Reutter, Werner

doi:10.1515/cclm.1988.26.9.559

Cited by 3 publications

(1 citation statement)

References 16 publications

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“…Despite several reports on the successful isolation of IgM antibodies by hydroxylapatite HPLC (HA-HPLC) or HA low pressure chromatography [3] , aggregation and precipitation of these antibodies during application to the column and at the beginning of the separation process present a serious problem, when only low salt concentrations are used. Similar problems are likely to occur, if IgM antibodies are separated by ionic exchange HPLC.…”

Section: Isolation Of Ig M Antibodies By Hydroxylapatite Hplcmentioning

confidence: 99%

Purification of Monoclonal Antibodies by Hydroxylapatite HPLC and Size Exclusion HPLC

Josić¹,

Löster²,

Kuhl³

et al. 1991

Biological Chemistry Hoppe-Seyler

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Monoclonal antibodies of both the IgG and the IgM type were purified by hydroxylapatite HPLC (HA-HPLC) under very mild conditions. The IgM type antibodies, which were isolated from ascites fluid and separated from other proteins also by means of size exclusion HPLC.It was shown that the most frequently observed disadvantage of HA-HPLC, that is the relative short life of the columns (P. Steifen (1989) GIT Fachz. Lab., Suppl. 3/89 (Chromatogr.}, 50-90), is due to microbial contamination rather than lower mechanical stability. In order to monitor column performance, a test was developed based on the use of standard proteins under isocratic separation conditions. This allows a direct comparison between the respective performances of columns made from different materials, hydroxylapatite or fluoroapatite, from different sources and with different particle sizes. A problem which often occurs with HA-HPLC in the case of IgM antibody isolation, namely precipitation of the antibodies at low salt concentrations at the beginning of a Chromatographie run, was avoided by adding sodium chloride to both separation buffers. Reinigung monoklonaler Antikörper durch HPLC an Hydroxylapatit und durch Size-Exclusions-HPLC

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Section: Isolation Of Ig M Antibodies By Hydroxylapatite Hplcmentioning

confidence: 99%

Purification of Monoclonal Antibodies by Hydroxylapatite HPLC and Size Exclusion HPLC

Josić¹,

Löster²,

Kuhl³

et al. 1991

Biological Chemistry Hoppe-Seyler

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Calcium-binding proteins 33 kDa, 35 kDa, and 65/67 kDa in normal rat and morris hepatoma tissues

et al. 1990

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Polyclonal antibodies were raised against membrane-associated calcium-binding proteins (apparent molecular masses 65000 and 67000 (CBP 65/67) and 33000 and 35000 (CBP 33 and CBP 35)), which were isolated from rat liver and Morris hepatoma. Using immunoblotting, various amounts of CBP 33 and CBP 35 as well as CBP 65/67 were detected in most rat organs. Using alkaline phosphatase and monoclonal-anti-alkaline phosphatase antibodies (APAAP), all the calcium-binding proteins were detected by immunohistochemical techniques in the plasma membranes of many cells, such as vascular endothelial cells, lymphocytes, epididymal principal cells, secretory and excretory duct cells of certain exocrine glands, straight distal tubular cells of the kidney, and in the cytoplasm of muscle cells and fibres as well as nerve cells and chondrocytes, and in connective tissue elements. Immunohistochemical analysis also showed that in polarized epithelial cells, e.g., renal tubular cells, epididymal principal cells or excretory duct cells, these calcium-binding proteins are present exclusively or mostly in the luminal plasma membrane.

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Receptor inhibition by immunoglobulins: Specific inhibition by autistic children, their relatives, and control subjects

Cook

Perry

Dawson

et al. 1993

J Autism Dev Disord

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Forty-two parents of children with autistic disorder, 15 children with autistic disorder, 17 siblings of children with autistic disorder, and 12 unrelated normal adult controls were studied to determine if immunoglobulins isolated from their plasma would inhibit binding of the 5HT1A agonist, [3H]-8-hydroxy-N,N-dipropyl-2-aminotetralin (DPAT) to 5HT1A receptors in human hippocampal membranes. There were no significant differences among the means of percentage inhibition of DPAT binding of parents, children with autistic disorder, siblings, or unrelated controls. In addition, there were no differences in the proportion of subjects with > 15% DPAT inhibition among autistic children, their parents, their siblings, or unrelated controls. Immunoglobulin inhibition was not specific for the 5HT1A receptor binding site, since immunoglobulins inhibited binding to 5HT2, D1, D2, and alpha 2-adrenergic binding sites. The immunoglobulins isolated from normal controls inhibited [3H]-rauwolscine binding at alpha 2-adrenergic sites less than immunoglobulins of children with autistic disorder and their parents and siblings. This study did not support the hypothesis that autoantibodies to 5HT1A or 5HT2 receptors are characteristic of autistic disorder.

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Isolation of Immunoglobulins and Their Use in Immunoaffmity HPLC

Cited by 3 publications

References 16 publications

Purification of Monoclonal Antibodies by Hydroxylapatite HPLC and Size Exclusion HPLC

Purification of Monoclonal Antibodies by Hydroxylapatite HPLC and Size Exclusion HPLC

Calcium-binding proteins 33 kDa, 35 kDa, and 65/67 kDa in normal rat and morris hepatoma tissues

Receptor inhibition by immunoglobulins: Specific inhibition by autistic children, their relatives, and control subjects

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