Development and Characterization of a Multiplex Bead-Based Immunoassay To Quantify Pneumococcal Capsular Polysaccharide-Specific Antibodies

Abstract

<jats:title>ABSTRACT</jats:title><jats:p>Enzyme-linked immunosorbent assay (ELISA), the traditional antibody quantification technique, has several limitations, especially when used to evaluate multivalent and/or infant vaccines. We have developed a multiplex bead-based antibody quantification assay (MBIA) to measure antibody response to multiple pneumococcal (Pn) serotypes (St) in a single assay. MBIA was compared with the WHO ELISA using a WHO panel of 12 international calibration sera for 7 Pn Sts. An agreement of 75 to 92% was obtained for all 7 Sts. MBIA exhibited good robustness, with the assay variability at ≤16%. A major contributor to MBIA variability was the cell wall polysaccharide (CWPs) content in Pn St-specific capsular Ps. This necessitated careful CWPs (20 μg/ml) preadsorption of sera. MBIA is specific, robust, and reproducible and offers high throughput. The use of MBIA will greatly reduce the cost and time required to evaluate the immune response to multiple Pn Sts and could help promote the licensure of future Pn and other multivalent vaccines.</jats:p>

Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle

“…In the case of multiplex bead-based immunoassays, comparison of the Luminex is frequently made with serological assays, such as ELISA. Correlation of these two methods varies among literature [ 34 , 54 – 59 ]. In the current study, agreement between the assays was generally good when the same (recombinant) antigen was used.…”

Maternal pneumococcal capsular IgG antibodies and transplacental transfer are low in South Asian HIV-infected mother-infant pairs

“…[16] A set of standard sera with known Spn IgG concentrations were used as controls in the assay. The tested serotypes represent 9 of the 13 serotypes included in the new conjugate pneumococcal vaccines that were developed for children below 3 years of age and for which our laboratory had assays readily available.…”

Development and Validation of 13-plex Luminex-Based Assay for Measuring Human Serum Antibodies to Streptococcus pneumoniae Capsular Polysaccharides

“…Therefore, the availability of methods that can be used with a reduced volume of serum sample is critical. Multiplexed fluorescent microsphere-based assays that have shown to be variable with respect to success in determination of pneumococcal serotype-specific IgG levels have been previously described ( 9 – 15 , 18 – 23 ).…”

“…In the WHO ELISA ( 8 ), each test yields one serotype-specific result per sample on a given assay plate, and adding tests to accommodate additional serotypes requires a significantly greater volume of serum, which is already limited, given the testing requirements of concomitantly administered pediatric vaccines. Microsphere-based Luminex immunoassays overcome this limitation by using spectrally distinct fluorescent microspheres as the solid support matrix onto which the target antigens are bound to simultaneously measure antibodies against multiple analytes from a single reaction well ( 9 – 15 ). For evaluation of humoral immune responses to pneumococcal vaccines, spectrally distinct fluorescent microspheres are coated with different PnPSs, which allows detection of multiple antigen-specific IgG antibodies simultaneously in a single reaction well, thus reducing analysis time and cost.…”