Abstract
Objective : Study goal was to investigate the nuclease like catalysis function of polyclonal IgM Abs recovered form rheumatoid arthritis (RA) patient sera. Methods : Intact polyclonal IgM Abs was recovered from the RA patient in a single step using CIM ® EDA monolith column. The DNA hydrolysis and RNA assays, in-situ and thermal inactivation, were performed to examine and measure the catalysis activity. Results: The RA IgM Abs exhibited moderate hydrolysis of plasmid DNA, pUC18 and total yeast RNA, respectively. Non-negligible differences in the nuclease activities were prominent among the patient’s samples, in spite of an overall moderate activity. Result of RNA in-situ assay revealed that the catalysis function is mediated by µ chain, thereby confirming it to be an intrinsic property of IgM Abs. Conclusion: The current preliminary study has concluded that IgM Abs from RA disease possess nuclease activity and IgMs mediate hydrolysis of both DNA and RNA macromolecules.
Highlights
The current preliminary study has concluded that IgM Abs from rheumatoid arthritis (RA) disease possess nuclease activity and IgMs mediate hydrolysis of both DNA and RNA macromolecules
A few of these antibodies (Abs), IgG class, are well-known to possess protease and nuclease-like enzymatic functions. Such Abs are designated as natural catalytic antibodies (CatAbs).[1]
Many research works have corroborated the presence of catalytic framework in the CatAbs and have revealed that it almost mimics the natural enzymes
Summary
Intact polyclonal IgM Abs were recovered from the sera, in a single step, using CIM® EDA monolith column. The DNase activity and RNA hydrolysis assays, viz. In-situ and thermal inactivation, were performed to achieve the objectives. Patient sera The RA serum (about 0.2 ml) samples of 20 cases, who fulfilled the American College of Rheumatology (ACR) criteria, were obtained from the biobank of Groupe hospitalier Cochin-Saint, Paris, France. Healthy serum (n=5) was obtained from the biobank. IgM purification Recovery of IgM Abs is explained in detail elsewhere.[11] In brief, purification of IgM Abs was carried out using CIM® Ethylenediamine (EDA) monolith column (BIA separation, Slovenia). About 50 μl of total serum, diluted twice in phosphate buffer pH 7.2, was injected into a disk type (dimension 12 X 3 mm; 0.34ml column volume) column.
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