Polymonoclonal (Not Polyclonal) Antibodies Derived from Convalescent Human B Cell Hybridomas Might Be a Better Therapeutic Option than Single Target Monoclonal Antibodies
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Both human B cell hybridoma technology and convalescent plasma therapy are promising immunological tools for therapeutic interventions. Here we propose using antibody producing B cells from convalescent SARS-CoV2 patients for developing human B cell hybridomas, and a combination of monoclonal antibodies against multiple immunogenic targets of SARS-CoV-2 spike protein might deliver an antibody cocktail for long-lasting therapeutic targeting.Keywords:
Polyclonal antibodies
Three enzyme immunoassays (EIA), two polyclonal and one monoclonal, and one polyclonal radioimmunoassay (RIA) for the detection of human immunodeficiency virus antigen (HIV-Ag) have been evaluated and compared. The three EIAs had similar sensitivity in detecting HIV viral lysate and were more sensitive than the RIA, which was the only assay with a one-step probing-detection format. However, the EIA that used a monoclonal anti-p24 as the capturing reagent was unable to detect any of the serial supernatant samples of a positive viral culture from an HIV-infected patient. Only the two polyclonal, non-p24-restricted assays were able to detect an unusual expression of HIV-Ag in the serum of an acute HIV-infected patient. Overall, the sensitivity of HIV-Ag capture assays was enhanced when (a) the capture antibody was polyclonal rather than monoclonal, (b) the polyclonal antibody was broad rather than p24-restricted, and (c) the probing-detecting procedure was in a two-step format rather than one-step format. In addition, the use of neutralizing assays to confirm the results was absolutely necessary.
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Weak expression of ABO antigens in newborns has been known for many years. In the near future reduced availability of polyclonal sera for ABO testing is to be expected. Therefore, it was the aim of our study to compare the suitability of monoclonal reagents for ABO testing of newborns to that of polyclonal sera.One monoclonal and 1 polyclonal reagent of each of the specificities anti-A, and anti-B and anti-AB from 7 manufacturers were tested by titration with blood from 5 newborns of blood group A, 5 of blood group B, and 3 of blood group AB.All monoclonal anti-A reagents showed better results than the polyclonal sera (for newborns of blood group A on the average of the 7 pairs of reagents 1.3 geometrical titration steps). Most of the monoclonal anti-B reagents showed higher titers than the polyclonal sera (on the average for newborns of blood group B 0.7 titration steps). Also most monoclonal anti-AB reagents were stronger than the polyclonal sera in testing newborns of blood group B the monoclonal anti-AB-reagents were slightly weaker than the polyclonal sera (on the average 0.4 titration steps).Use of monoclonal reagents for ABO testing of newborns shows some advantages in comparison to the use of polyclonal sera.
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Results obtained measuring blood Cyclosporine A (CsA) concentrations in transplanted patients (124 samples of cardiac, 20 samples of liver, and 10 samples of kidney transplanted patients) by the use of two monoclonal radioimmunoassay (RIA) methods have been compared with those found using the HPLC technique (considered as the reference method) and two polyclonal RIAs. In addition, results on quality control samples collected in a multicentre collaborative study for CsA assay from the users of the same monoclonal and polyclonal RIAs were analysed to evaluate the performance of the methods under study. Polyclonal RIAs, which measure both the parent molecule and its metabolites, produced results 1.5-3 times higher than HPLC or monoclonal RIAs. On the contrary the two RIAs, which use monoclonal antibodies specific for CsA, show a better correlation with HPLC; these RIAs, which measure the intact drug molecule only, are recommended when the monitoring of the native molecule of CsA is requested. As far as the reproducibility is concerned, the four RIAs (both polyclonal and monoclonal) exhibit an unsatisfactory degree of between-assay and between-lab precision, since the coefficients of variation (CVs) ranged from 19.4% to 23.1%.
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To develop and evaluate an enzyme-linked immunosorbent assay (ELISA) for detecting adenovirus antigens in which a group-specific monoclonal antibody to the adenovirus hexon is used, we studied two different ELISA test systems. The test system which was the most sensitive and specific was then compared in parallel tests with a similarly constructed ELISA in which a mouse polyclonal serum was used. Both the ELISA with the monoclonal antibody and that with the polyclonal serum detected purified hexon and 15 different adenovirus types with similar sensitivities. The two assays also showed no reaction with 23 heterologous viruses. Both tests detected adenovirus in stool and respiratory tract specimens tested for adenovirus by standard tissue culture techniques with similar sensitivities and specificities, but neither was sufficiently sensitive for routine testing of these types of clinical specimens. However, the ELISA with the monoclonal antibody proved to be a good test for the noncultivatable adenoviruses, detecting 12 of 12 stool samples that were positive by electron microscopy. The monoclonal antibody proved to be as sensitive and specific as the polyclonal serum and has the advantage that it can be produced in unlimited quantities and needs to be characterized only once.
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Mastadenovirus
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Monoclonal proliferative lesions may be identified by X chromosome inactivation skewing relative to normal polyclonal tissues. We have quantitatively analyzed X-inactivation patterns throughout polyclonal uterine tissues to develop interpretive criteria for recognition of monoclonal neoplasms. Six fresh tissue samples (two samples each of cervix, endometrium, and myometrium) were collected from hysterectomy specimens, and the percentage of androgen receptor (HUMARA) marker allele present on inactive X chromosomes was calculated from a PCR assay. Exact balancing yields 50% of the marker on the inactive X, whereas complete skewing shows either 0 or 100%. X inactivation was similar throughout the tissues of each uterus but was significantly different among the 11 women studied. Comparison of differences in X inactivation between pairs of polyclonal tissue samples within each uterus (Xi spread) permitted delineation of cumulative experimental and biological variation of this parameter. Polyclonal-polyclonal Xi spread averaged 10.7 and was independent of the tissue type, sampling site, or the individual studied. Severe baseline skewing of reference polyclonal tissues or contamination of monoclonal tissue by polyclonal cells may reduce the polyclonal-monoclonal Xi spread. The extent of X-inactivation skewing necessary to infer a monoclonal process should exceed the 20 or 27 point spread seen, respectively, between 85 and 95% of polyclonal samples.
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Abstract The electrophoretic pattern of rheumatoid factor (RF) was investigated in 40 polyclonal sera, by using radiolabeled IgG. Thirty sera specifically bound IgG aggregates, correlating with their RF titer. The binding pattern was monoclonal or oligoclonal. The molecules responsible were classic RF antibodies, as shown by using purified IgM, inhibition experiments, and by the optimal size of aggregates (2,000–4,000 kd). These data show that RF heterogeneity is restricted in polyclonal sera, and this can have a bearing on several mechanisms.
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Abstract We have prepared a monoclonal antibody to Lipoprotein(a)[Lp(a)] and have used it to develop an ELISA test for assaying Lp(a) in serum. The monoclonal antibody employed in the assay system reacts uniformly with S1,S2,S3 and B phenotypes of isoforms, and no cross‐reaction with plasminogen at a concentration of 100 mg/dL was observed. Results of the monoclonal ELISA assay were similar to those obtained with a polyclonal antibody ELISA method and demonstrated a correlation coefficient, r=0.99 with the equation for the regression line: Y(proposed)= 1.06 X (polyclonal antibody reference ELISA test) = 0.36(N = 51). Inter‐ and intra‐assay precision(CVs) of the monoclonal ELISA assay were between 2.2–3.6% at a mean Lp(a) concentration range of 19.1–68.2 mg/dL,(N = 12). Assay results of various standards were compared by both monoclonal and polyclonal antibody ELISA tests. We observed some discrepancies between expected concentrations and the polyclonal antibody ELISA assay results, which is thought to be more uniformly reactive to the various Lp(a) phenotypes. The monoclonal antibody employed in our proposed method reacts uniformly with Lp(a) phenotypes, and the assay exhibits excellent sensitivity, specificity, and accuracy and is well suited for clinical use.©1995 wiley‐Liss, inc.
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The use of monoclonal antibodies (mabs) to blood group antigens is constantly increasing for routine typing. Two heterohybridoma cell lines, HMR15 and HMR22, were established by Epstein-Barr virus transformation of peripheral blood lymphocytes from a blood donor with anti-Di a . HMR15 mab directly agglutinated Di(a+) red cells, and HMR22 mab agglutinated Di(a+) red cells exclusively by the indirect antiglobulin test. Reactivities of both HMR15 and HMR22 mabs were specific for Di a and had good correlation with the reactivity of a commercial, polyclonal antiserum. The binding of monoclonal antibodies to antigen-positive red cells was mutually blocked by each other as well as by polyclonal anti-Di a . Immunoprecipitates by HMR22 mab with a Di(a+) preparation showed a 120kDa band that was stained by anti-band 3. Di a typing of 2427 blood donors with the mabs and polyclonal typing serum detected 244 Di(a+) individuals (10.1%). No discrepancy was observed between the mabs and the polyclonal antiserum. HMR15 and HMR22 mabs are useful Di a typing reagents and can substitute for commercial antiserum. Immunohematology 2000;16:78–81.
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We report the cross-reactivities and comparative molecular masses of the IgE ε chains in humans, rats, mice, and dogs. Monoclonal human, rat, and mouse IgE, and our purified polyclonal dog IgE were used in the study. IgE of the 4 species, separated by SDS-PAGE, were analyzed by immunoblotting with polyclonal and monoclonal antihuman IgE, polyclonal and monoclonal antimouse IgE, monoclonal antirat IgE, and polyclonal antidog IgE antibodies. The polyclonal anti-human and polyclonal antimouse IgE cross-reacted with the IgE of the other 3 species, while their monoclonal forms cross-reacted with dog IgE only. Polyclonal antidog IgE cross-reacted with human and mouse IgE, while the monoclonal antirat IgE did not cross-react with any other species. 'Reverse' passive cutaneous anaphylaxis in ragweed-sensitized dogs revealed that polyclonal anti-human and polyclonal antimouse IgE were able to elicit positive skin responses, and monoclonal antihuman, antirat, and antimouse IgE antibodies were not. The molecular masses of the ε chains were 77 kDa for mice, 75 kDa for rats and dogs, and 70 kDa for humans.
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