A Quantitative Immunochemical Measure of thePrimary Interaction Between I*BSA and Antibody

Farr, Richard S.

doi:10.1093/infdis/103.3.239

Cited by 896 publications

(213 citation statements)

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“…The RIPA technique was modified to estimate values for the binding affinity of MAbs to virus particles. In this method, modelled on that of Farr (1958), 100 ~tl volumes of each MAb, diluted to the previously determined 50 % endpoint, were mixed with 10 ~tl volumes from a range of radiolabelled virus dilutions, of known antigen concentration, starting at 5000 c.p.m. Subsequent stages followed the procedure described for RIPA except that antimouse antibody was added to all reaction mixtures.…”

Section: Radioimmunoprecipitation Assays (Ripa)mentioning

confidence: 99%

Characterization of monoclonal antibodies raised against a synthetic peptide capable of inducing a neutralizing response to human rhinovirus type 2

Barnett

Rowlands

Parry

1993

Journal of General Virology

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Synthetic peptides incorporating the derived amino acid sequence of VP2 residues 156 to 170 of human rhinovirus type 2 (HRV2) have previously been shown to elicit antibodies that neutralize virus infectivity. The proportion of virus-reactive antibodies present in polyclonal antisera to these peptides is, however, very low. Moreover, neutralization titrations of such antisera correlate poorly with other assays of either anti-virus or anti-peptide activity, suggesting the presence of antibodies with different specificities. To investigate these findings further, we produced a panel of monoclonal antibodies (MAbs) to VP2 peptides of residues 156 to 170 and characterized their reactions with a range of antigens in ELISA, precipitation and neutralization titrations. All the MAbs obtained recognized the homologous peptide, but could be divided into four main reaction groups according to their specificity for viral antigens. Antibodies in the first group recognized and neutralized native virus, apparently by preventing attachment to cells. A second group of MAbs bound to intact particles with similar affinities to the first group, but failed to neutralize infectivity. Antibodies in the third group recognized virus only after capsid distortions incurred by heating or by previous reaction with polyclonal antibodies. The fourth group comprised MAbs that were mainly peptide-specific. Some possible applications of anti-peptide MAbs to improving the design of peptide immunogens are considered.

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Section: Radioimmunoprecipitation Assays (Ripa)mentioning

confidence: 99%

Characterization of monoclonal antibodies raised against a synthetic peptide capable of inducing a neutralizing response to human rhinovirus type 2

Barnett

Rowlands

Parry

1993

Journal of General Virology

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“…The number of plaque-forming cells (PFC) in the draining lymph nodes and the serum antibody levels were measured [10][11]18, and 30-33 days after DNP-GPA immunization. In addition, the avidity of antibody secreted by PFC was measured by a hapten-inhibition technique which allows the enumeration of PFC producing antibody of any given avidity (14).…”

mentioning

confidence: 99%

Hapten-Specific Tolerance

Davie

Paul

Katz

et al. 1972

The Journal of Experimental Medicine

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The treatment of guinea pigs with 2,4-dinitrophenyl (DNP) t derivatives of the copolymer of D-glutamic acid and D-lysine (D-GL) induces a state of specific immunologic tolerance in the recipient (1). The type of tolerance induced is unusual in that it is DNP specific. That is, challenge of such guinea pigs with a DNP conjugate of ovalbumin (DNP-OVA) elicits anti-DNP responses markedly smaller than those of animals which have not been pretreated with DNP-B-GL. The degree of depression in responsiveness is related to the dose of tolerogen, to the mode of immunization with DNP-OVA, and to the interval between the two events. A similar type of haptenspecific tolerance, in mice, has recently been described by both Hraba et al. (2) and by Golan and Borel (3).The tolerant state, in this model, appears to be largely expressed in the precursors of antibody-secreting cells. The evidence for this view is the following:(a) The number of DNP-specific antigen-binding lymphocytes in the lymph nodes of tolerized animals is markedly reduced (1). These antigen-binding lymphocytes are members of the bone marrow-derived (B) lymphocyte pool and include the precursors of antibody-secreting cells (4).(b) The tolerance is hapten specific (1). Thymus-derived (T) lymphocytes acting as "helpers" in the activation of precursor cells are almost exclusively carrier or conjugate specific (5, 6); on the other hand, hapten-specific precursors of antibodysecreting cells are relatively common (4, 7). Hapten-specific tolerance thus implies precursor cell tolerance.(c) Even when an excess of helper cells are provided through an immunization with carrier after tolerance induction with DNP-D-GL but before challenge with DNPcarrier conjugate, the subsequent anti-DNP response is markedly diminished (1). Thus, in a situation in which helper T cells are not limiting, the hapten-specific tolerance is easily demonstrable, again implying a defect in the population of precursors of antibody-secreting cells.

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“…The PEG-treated cells were seeded into 24-well Costar plates, and hybrids were selected in growth medium containing 100 PM hypoxanthine, 0.4 PM aminopterin, and 16 I.IM thymidine (HAT medium) [19]. Hybrids producing mouse immunoglobulins were screened by radioimmunoassay (RIA) [ 16,171 and grown in the culture medium for the classification of mouse immunoglobulins or were injected intraperitoneally into mice for further growth and production of antibody in the ascites fluid. The cell culture or ascites fluids were used for analysis of antibody production by Ouchterlony immunodiffusion and radioimmunoassay and by their effects on enzyme activity.…”

Section: Methodsmentioning

confidence: 99%