VERSATILITY OF ELISA (ENZYME-LINKED IMMUNOSORBANT ASSAY)

MarinBio offers ELISA as a highly quantitative and sensitive immunobioassay method for detecting and quantifying antibodies, cytokines, hormones, proteins and other biologic drugs as well as DNA for which there is a specific antibody. Sensitivities for the ELISA can be in the pg/mL range. The MarinBio scientists have developed ELISAs from inception of which the analyte is specifically detected in a complex matrix, such as blood, plasma, serum from human or various species or in cell culture media. The MarinBio scientists have developed a 90 min GMP ELISA and manufactured several thousand GMP ELISA kits, distributed to hospital med-tech laboratories and trained the technicians. It was used to differentiate appropriate clinical treatment.

MarinBio clients has successfully completed ELISA service work for the following common examples:

Pre-clinical and clinical PharmacoKinetic (PK) studies of amounts of drug present after administration

Mechanism of Action (MOA) of a drug

Host Anti-Drug Antibodies (ADA) as a measure of immunogenicity

Production of hormones or inflammatory cytokines

Phosphorylated proteins as part of a signaling pathway

Drug release assays

IND application requiring GLP or GMP level

Drug screening or discovery at the research level (non-GLP/GMP)

MarinBio scientists has significant experience with direct or competitive ELISA assays. Additionally, we develop ELISAs using adherent cells or membranes to measure analyte concentration. Our experience includes homogeneous ELISAs and sandwich ELISAs with very large analytes (~800,000 daltons) or with small mimetics or peptides. Most clients request direct or competition ELISAs with colorimetric, fluorescent, luminescent and time-resolved readouts. We have ability to measure high-throughput ELISAs and multiplex ELISA bioassays for multiple analytes in a single sample. Also our scientist can perform ELISpot (Enzyme-Linked ImmunoSpot) for very sensitive single cell cytokine production.

ELISA has been a workhorse method at MarinBio for the past 25 years. MarinBio scientists have deep experience with off-the-shelf and custom developed ELISA assays, as well as ELISA qualifications and validations.

Top Figure Legend. The ELISA shown measures the increased production of IL-10 in response to increasing drug concentration found in the supernatant of cells incubated with either Reference Standard or Test Sample measured by ELISA.

Bottom Figure Legend. Correlation of ELISA (sensitivity 2pg/mL) and ELISPOT for measuring the gamma-IFN response of peripheral blood T-cells to a specific CytoMegaloVirus (CMV) oligopeptide. The CMV oligopeptide was tested at 10-fold dilutions from 10-9 M to 10-15 M.

In summary, ELISA is a highly quantitative and sensitive bioassay that is conveniently automated for microwell plate-based assays (including high-throughput and multiplexed ELISA) for detecting and quantifying antibodies, cytokines, hormones, proteins, oligopeptides and other specific antigens, including pharma drug antigens. ELISA has exquisite sensitivity and uses a very highly specific antibody coupled with an enzyme to quantitate the analyte.