The role of pH in modified ELISA procedures used for the estimation of functional antibody affinity

Green, David; Etten, Lisette van; Milligen, F.J. van; Aalberse, Rob C.; Turner, Malcolm

doi:10.1016/0022-1759(93)90369-i

Cited by 38 publications

(14 citation statements)

References 9 publications

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“…The pH of the high-concentration solution was reduced to 6.75, compared to pH 7.4 for PBS alone. However, Goldblatt et al (13) showed that pH has no effect on the thiocyanate ion. Although antibody avidity is an important surrogate of protective efficacy for several vaccine types, vaccines differ in the ability to evoke avidity maturation.…”

Section: Discussionmentioning

confidence: 99%

Antibody Avidity and Immunoglobulin G Isotype Distribution following Immunization with a Monovalent Meningococcal B Outer Membrane Vesicle Vaccine

et al. 2002

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Section: Discussionmentioning

confidence: 99%

Antibody Avidity and Immunoglobulin G Isotype Distribution following Immunization with a Monovalent Meningococcal B Outer Membrane Vesicle Vaccine

et al. 2002

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“…Avidity of MenC-specific IgG was measured by an ELISA as described above, including a potassium thiocyanate (KSCN) elution step (16). After the serum incubation step, serial dilutions of KSCN (7.5 to 0.117 M) or PBS-Tween (100% binding) were added to the wells and incubated for 15 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Concomitant Administration of Mycobacterium bovis BCG with the Meningococcal C Conjugate Vaccine to Neonatal Mice Enhances Antibody Response and Protective Efficacy

Brynjólfsson

Bjarnarson

Mori

et al. 2011

Clin Vaccine Immunol

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Mycobacterium bovis BCG is administered to human neonates in many countries worldwide. The objective of the study was to assess if BCG could act as an adjuvant for polysaccharide-protein conjugate vaccines in newborns and thereby induce protective immunity against encapsulated bacteria in early infancy when susceptibility is high. We assessed whether BCG could enhance immune responses to a meningococcal C (

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“…Serotype-specific antibody avidity indexes (AIs) for sera and MAbs were evaluated using a modification of a standard ELISA in which an incubation step using the chaotropic agent ammonium thiocyanate (NH 4 SCN) was incorporated into the assay. The method has been described in detail previously (16), standardized for Hib and meningococcus C (15,17), and adapted for pneumococcal antibody avidity determination. In brief, antibodies were diluted in BSA-PBS-T to a concentration known to give an optical density of 1 in the standard pneumococcal ELISA, plated out onto appropriate serotype-coated plates, and incubated for 2 hours.…”

Section: Methodsmentioning

confidence: 99%

Correlation of Molecular Characteristics, Isotype, and In Vitro Functional Activity of Human Antipneumococcal Monoclonal Antibodies

Baxendale¹

2006

Infect Immun

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Structure-function correlations of pneumococcal antibodies are important in predicting how changes in the pneumococcus (Pnc)-specific B-cell repertoire will influence humoral immunity against invasive Pnc disease. Using a unique panel of human hybridomas derived from memory B cells after pneumococcal conjugate vaccination, we analyzed the structure-function relationship of nine monoclonal antibodies (MAbs) reactive to Pnc polysaccharides. The avidities of the antibodies correlated with the avidity of donor immune serum (R, 0.7; P < 0.025), and this relationship was particularly strong for immunoglobulin A clones (R, 1; P < 0.0005), suggesting that the MAbs may represent important clones contributing to serological memory. Common heavy-light chain combinations and amino acid replacement mutations were seen for clones with the same serospecificity from different individuals. The two highest-avidity MAbs used Vh3-48, and two MAbs with the same serospecificity, using the same V gene pairings (Vh3-7 and Vk2A17), had similar avidities, suggesting that canonical V gene use makes an important contribution to avidity. Although all clones had mutation levels consistent with their being derived from memory B cells, low levels of replacement mutation were associated with high avidities. This relationship was strongest for Vh genes (R, 0.8; P < 0.01). Opsonophagocytosis was demonstrated for all clones, and there was a trend toward clones using canonical genes with low levels of mutation having high opsonophagocytic activities (R, 0.5). These data suggest that the use of canonical genes in the Pnc antibody response is associated with highly functional antibodies and that most somatic mutations seen in these genes are not antigen selected.Understanding the molecular diversity of the antibody response to polysaccharide antigens has assumed increasing importance with the development of protein-polysaccharide conjugate vaccines that are immunogenic and protect those who are unable to respond to pure polysaccharide vaccines. Many studies have focused on responses to the capsular polysaccharides derived from Haemophilus influenzae type b (Hib), and these studies have demonstrated an oligoclonal antibody (Ab) response with between one and four immunoglobulin (Ig) variable-region (V) genes dominating the Ab repertoire to a specific polysaccharide (2, 25). Similar heavy-chain variable-region genes have been shown to be used by different individuals, and particular V genes appear to be associated with superior functional activity in vitro (24,26). A number of factors have been shown to influence antigen-specific V gene dominance, including the age of the individual (3), the dose of antigen (18), and the vaccine formulation (24). It is now becoming clear that the choice of V-region gene during a primary immune response may be critical, as this Vregion gene use may dominate subsequent memory responses. A murine study by Sanchez et al. demonstrated that the V gene repertoire induced during the initial priming stage of a T-dependent antige...

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The role of pH in modified ELISA procedures used for the estimation of functional antibody affinity

Cited by 38 publications

References 9 publications

Antibody Avidity and Immunoglobulin G Isotype Distribution following Immunization with a Monovalent Meningococcal B Outer Membrane Vesicle Vaccine

Antibody Avidity and Immunoglobulin G Isotype Distribution following Immunization with a Monovalent Meningococcal B Outer Membrane Vesicle Vaccine

Concomitant Administration of Mycobacterium bovis BCG with the Meningococcal C Conjugate Vaccine to Neonatal Mice Enhances Antibody Response and Protective Efficacy

Correlation of Molecular Characteristics, Isotype, and In Vitro Functional Activity of Human Antipneumococcal Monoclonal Antibodies

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