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Corette Guitreau
We are thrilled to collaborate with your team and achieve our shared goals for your ELISA Assay Development and Immunoassay Validation. We can develop and validate de novo ELISA PK/ADA or ELISA biomarker assays in six to eight weeks. Then, we can deliver your audited report with detailed documentation of ELISA Immmoassay Validation experiments in a few weeks. We constantly stretch your budget by combining our scientific experience, regulatory expertise, and operational excellence.
We are eager to work with you on your ELISA Immunoassay project. We believe close communication is vital to success, and want to understand your needs and expectations in advance as much as possible. To help us prepare a detailed quote and timeline, please share additional information about your analyte(s), critical reagents and supplies, current and previous Enzyme Linked Immunoassay, study protocols, and sample sizes. Thank you for choosing us as your bioanalytical partner in cutting-edge science!
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Enzyme Linked Immunosorbent Assay (ELISA) method is a widely utilized immunoassay for quantitating and detecting proteins, hormones, peptides(through ELISA peptide), cytokines, antibodies, and other drugs along with their metabolites. ELISA methods are effective in quantitatively detecting any molecule, or antigen, that can be ascertained by an antibody. For example, ELISA methods are used in pregnancy testing, infectious disease identification, and detection of cytokines, and soluble receptor proteins, etc. Due to the precision, sensitivity, assay speed, and ease of quantitation, ELISA method development is a common choice for several diagnostic and research applications.
While there are various formats of ELISA method, the most common is a sandwich ELISA method. In this format, the analyte whose concentration is to be measured is sandwiched in-between two antibodies that bind to a different region (epitopes) on the antigen. These antibodies are referred to as detection antibody and capture antibody. In this assay, the capture antibody is coated to a microtiter plate in a 96-well format. The antigen binds to the capture antibody, and a detection antibody is used to measure the analyte. The antibody is conjugated to an enzyme, typically horseradish peroxidase (HRP), and detected via catalysis of a substrate which yields a colored product. Spectrophotometry is used to monitor the colored product, and a standard curve is utilized for calculating the antigen concentration in the sample. When the antigen is small, or two antibodies that can simultaneously bind to the antigen are unavailable, then another variation of ELISA, termed competitive ELISA method is utilized using a single epitope.
It is notable that the ELISA method typically uses 96 microwell plates for parallel analysis of many samples, standards, and controls in a single experiment. The surface of these plates is treated with special absorbents such that the antigen or antibody can adhere properly. ELISA method offers increased sensitivity and specificity compared to other standard antibody-based assays. In ELISA method, the interactions with analyte or antibody occur through serial binding that is done to some solid surface such as polystyrene in the 96 microwell plates discussed above.
Whenever an ELISA method is developed for any analyte, a surface attachment strategy is established first. Once the analyte is affixed to the surface of the plate, it is immobilized and helps with the sequential addition of other reagents and washing cycles in the next steps, without mixing. ELISA method is thus a solid-phase assay.
The process of immobilizing an antibody or antigen to the plate is essential, as the method of immobilization utilized is what determines the efficacy of the ELISA method. Many times, the immobilization may cause a conformational change in the analyte or antibody, preventing the detection antibody from binding. Thus, it is critical to immobilize the antibody or antigen while preserving its conformational structure.
The specificity and sensitivity of the assay are determined using a range of concentrations of antibody and/or antigen. The affinity of the detection antibody and sensitivity of the detection reagents corresponding to a specific antigen are principally responsible for defining the performance of an optimized ELISA method. Selection of the best capture and detection antibody is critical for effective assay development.
The antibodies which are labeled with an enzyme (detection antibodies) contribute to the signal output of this assay. Different types of enzymes can be utilized, such as alkaline phosphatase, horseradish peroxidase, etc., each of which produces a color upon reacting with their substrates. These colored products are detected with spectrophotometry, where the signal intensity is proportional to the amount of antigen in the microwell. Afterward, a standard curve along with positive and negative controls is used to quantify the amount of analyte within a given sample.
Proper ELISA method development and validation are critical for the accurate quantification of an analyte. NorthEast BioLab helps you select the right ELISA method format and develop a robust protocol for analysis. Our specialists optimize the assay by titrating various components to ensure accurate results.
Our pharmacokinetic studies for quantitation of drug-responsive levels of a specific serum protein using ELISA protein assay development allow accurate and reliable pharmacokinetic data to help regulate dosage regimens.
As in pharmacokinetic studies, an ELISA method can be used in toxicokinetic studies to quantitate a drug or compound when administered at high dosage. TK studies allow estimation of the level of toxicity that can be produced by these drug compounds.
ELISA method is hugely popular as a fundamental, cost-effective technique that offers high precision and broad dynamic range in addition to specificity, selectivity, and reliability for biomarker testing and quantitation.
An ELISA method can be used for immunogenicity testing. These studies are carried out to assess how drug exposure induces an immune response in the body of humans or other animals. It further evaluates how anti-drug antibodies (ADA) can affect immunogenicity.
Our scientists help clients with the successful execution of cell-based assays to assess the toxicity of the compounds and ensure reliable drug manufacturing. For example, we perform cytotoxicity testing and mechanism of action (MOA) assay to understand the biochemical reactions triggered by drug compounds.
At NorthEast BioLab, we ensure robust ELISA methods to quantify large molecule analytes such as biologics, proteins, and antibodies in your samples. As often as it takes, our veteran scientists with 30+ years experience discuss and study your ELISA method development and validation in depth. Our team continues without yielding until we have customized these assays per your requirement and satisfaction.
We encourage intensive collaboration between the client and our scientists for complete transparency throughout your study with us. NorthEast BioLab offers quick turnaround times, scalability, and flexibility, given our extensive experience with ELISA method development and a thorough understanding of the science behind it. We enable our clients to expedite drug approval through efficient assay development, validation, and optimization while maintaining 100% regulatory compliance.
ELISA Method, based on colorimetric or chemiluminescent reaction, is the best-in-class ligand binding assay (LBA) predominantly used for large molecule bioanalytical method development and sample analysis. Scientists and regulatory agencies widely trust the direct, indirect, sandwich, and competitive variants of ELISA Development and Method Validation for diverse PK, BA/BE, TK, Immunogenicity (ADA), and Biomarker testing. Our veteran team undertakes Enzyme Linked Immunosorbent Assay development from scratch using available antibodies, diluents, buffers, substrates, reagents, control matrices, as well as research use only (RUO) kits. NorthEast BioLab delivers research mode, and fully GLP validated ELISA Method and Assay Development at an unmatched turnaround and value to sponsors.
ELISA Method is an abbreviation for enzyme-linked immunosorbent assay, also called enzyme immunoassays (EIA). The fundamental principle used in ELISA method development is the use of antibodies to detect and quantify proteins, peptides (through ELISA peptide), hormones, or other small molecules in preclinical or clinical samples. In an ELISA method development, antigens are immobilized on solid support either directly or via the use of a capture antibody immobilized on the surface. Subsequently, we add a primary detection antibody. If the detection antibody binds to the analyte of interest, the complex can be directly detected and quantified by the chromogenic tag or a secondary enzyme-conjugated antibody. In our GLP ELISA lab, we typically perform method development, validation, and sample analysis in multi-well plates (96- or 384-wells) for capture antigen binding. This characteristic makes the ELISA assay easy to perform on multiple samples simultaneously with excellent selectivity.
ELISA immunoassays include a direct, indirect, sandwich, and competitive variants. In the direct ELISA method, an antigen is immobilized to the surface of a multi-well plate and detected with an antigen-specific antibody directly conjugated to HRP or another detection tag.
For an indirect ELISA assay, as with the direct ELISA method, an antigen is immobilized to the surface of the multi-well plate, but we need a two-step process for detection. The primary antibody binds to the antigen of interest, and then a secondary antibody labeled with a detection reagent is used for quantification. The sandwich ELISA method, the most widely used format, requires an antibody pair where each antibody is specific to a different epitope of the antigen. The first antibody, often called the capture antibody, is coated on the 96 well plate and binds to the analyte of interest. Once the unbound materials in the sample matrix get washed away, the second antibody conjugated with a detection tag quantitates the analyte. Competitive ELISA methods, often referred to as an inhibition ELISA assay or competitive immunoassays, measure the concentration of an antigen by detecting signal interference. We can adapt direct, indirect, or sandwich ELISA assays to the competitive format. Here, the antigen in the sample competes with a reference antigen pre-coated on a multi-well plate or added in the same plate wells. Consequently, lower concentrations of analyte produce a higher level of the signal. Some competitive ELISA assay, however, utilize a slightly different approach. These kits substitute a labeled antigen for the labeled antibody, but the principle of the ELISA protein assay remains the same.
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