Isolation of specific IgM monoclonal antibodies by affinity chromatography using alkaline buffers

Pl, Lim

doi:10.1016/0161-5890(87)90106-4

Cited by 29 publications

(2 citation statements)

References 11 publications

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“…It is more difficult to explain the ELISA profile (nil-IgM, high IgG) in Group D. One possibility is that the IgM antibodies, which are more fragile than IgG antibodies [20], are denatured due to storage of the specimen. Alternatively, in the case where both the 1 st and 2 nd specimens share a similar ELISA profile, this may be related to antibody production.…”

Section: Discussionmentioning

confidence: 99%

Combined Rapid (TUBEX) Test for Typhoid-Paratyphoid A Fever Based on Strong Anti-O12 Response: Design and Critical Assessment of Sensitivity

Yan

Tam²,

Kan

et al. 2011

PLoS ONE

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Rapid diagnostics can be accurate but, often, those based on antibody detection for infectious diseases are unwittingly underrated for various reasons. Herein, we described the development of a combined rapid test for two clinically-indistinguishable bacterial diseases, typhoid and paratyphoid A fever, the latter fast emerging as a global threat. By using monoclonal antibodies (mAbs) to bacterial antigens of known chemical structures as probes, we were able to dissect the antibody response in patients at the level of monosaccharides. Thus, a mAb specific for a common lipopolysaccharide antigen (O12) found in both the causative organisms was employed to semi-quantify the amounts of anti-O12 antibodies present in both types of patients in an epitope-inhibition particle-based (TUBEX) immunoassay. This colorimetric assay detected not only anti-O12 antibodies that were abundantly produced, but also, by steric hindrance, antibodies to an adjoining epitope (O9 or O2 in the typhoid or paratyphoid bacillus, respectively). Sensitivity and, particularly, reaction intensities, were significantly better than those obtained using an anti-O9 or anti-O2 mAb-probe in the examination of paired sera from 22 culture-confirmed typhoid patients (sensitivity, 81.8% vs 75.0%) or single sera from 36 culture-confirmed paratyphoid patients (52.8% vs 28.6), respectively. Importantly, sensitivity was better (97.1% for typhoid, 75.0% for paratyphoid) if allowance was made for the absence of relevant antibodies in certain specimens as determined by an independent, objective assay (ELISA) — such specimens might have been storage-denatured (especially the older paratyphoid samples) or procured from non-responders. Benchmarking against ELISA, which revealed high concordance between the two tests, was useful and more appropriate than comparing with culture methods as traditionally done, since antibody tests and culture target slightly different stages of these diseases. Paired sera analysis was insightful, revealing 64% of typhoid patients who had no change in antibody titer over 4–16 days, and 14% with no IgM-IgG class-switching.

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Section: Discussionmentioning

confidence: 99%

Combined Rapid (TUBEX) Test for Typhoid-Paratyphoid A Fever Based on Strong Anti-O12 Response: Design and Critical Assessment of Sensitivity

Yan

Tam²,

Kan

et al. 2011

PLoS ONE

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“…Otherwise, there might be certain degree of degradation of lg during our attempt to concentrate these proteins from the dilute column fractions. Lim (1987) has also not been able to concentrate the eluted proteins from weak solutions (< l g/ml), without gross denaturation. Further studies are being continued in this aspect to eliminate the anomaly and to purify the lg to homogeneity.…”

Section: •• Imentioning

confidence: 97%

Purification of Rohu Serum Immunoglobulin: A Preliminary Study

Mohanty,

Sahoo,

Mukherjee

2023

joa

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Attempts were made to purify immunoglobulins from serum of rohu, Labeo rohita, which had been immunised with Edwardsiella tarda. Initially, the proteins in the serum were salted out at 50% saturation with ammonium sulphate and were chromatographed successively by gel filtration and ion exchange columns. The E. tarda agglutination-positive fractions from ion exchange column when concentrated and checked in non-reduced SDS-PAGE, three bands were observed. Since teleost immunoglobulins have been shown to belong to a single class, the two extra bands found in our study might be the degradation products of immunoglobulin or some unpurified contaminants.

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Common occurrence of an antiidiotypic antibody that recognizes T14+ anti‐DNA antibodies in patients with systemic lupus erythematosus

Lim

Leung

et al. 1996

Arthritis & Rheumatism

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Objective. To investigate whether antibodies to a T14 anti-DNA antibody can be found in patients with systemic lupus erythematosus (SLE).Methods. Seventy-six serum samples (37 from patients with SLE) were randomly selected from among sera submitted for routine antinuclear antibody testing. Short, overlapping peptides based on the partial V, (variable region of the heavy chain) sequence of the T14 antibody were synthesized on multipins and screened for reactivity with SLE sera. In addition, selected peptides from T14 and related proteins were synthesized in bulk and screened for reactivity with both SLE and control sera. A monoclonal antibody was generated to determine the prevalence of the T14 idiotype (T14+ Id) in the different study populations.Results. Antibodies were detected by a peptide based on the third complementarity-determining region (CDR3) of the T14 protein in 15 (41%) of 37 patients with SLE or 15 (54%) of 28 who had anti-DNA antibodies, in 3 (9%) of 34 patients without anti-DNA antibodies (9 of whom had SLE), and in 6 (10%) of 57 healthy controls. In SLE sera, the antiidiotypic (anti-Id) responses (IgM and IgG) correlated well with the anti-DNA responses (IgG), and both responses correlated well with the T14+ Id activity in SLE sera. Control peptides based on the 18/2 (16/6+ Id) and S107 proteins detected low antibody activities in SLE sera, attributable to cross-reactivity with the T14 peptide. A peptide based on an unrelated human antibody was not reactive with these sera.Conclusion. Anti-Id antibodies directed to T14 V,CDR3 were found commonly in the sera of patients with SLE, and they appeared to be induced by the anti-DNA antibodies present in the sera. Based on these findings, these secondary antibodies may be pathogenic in SLE.

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Isolation of specific IgM monoclonal antibodies by affinity chromatography using alkaline buffers

Cited by 29 publications

References 11 publications

Combined Rapid (TUBEX) Test for Typhoid-Paratyphoid A Fever Based on Strong Anti-O12 Response: Design and Critical Assessment of Sensitivity

Combined Rapid (TUBEX) Test for Typhoid-Paratyphoid A Fever Based on Strong Anti-O12 Response: Design and Critical Assessment of Sensitivity

Purification of Rohu Serum Immunoglobulin: A Preliminary Study

Common occurrence of an antiidiotypic antibody that recognizes T14+ anti‐DNA antibodies in patients with systemic lupus erythematosus

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