elisa laboratory procedures
  • Troubleshooting Guide for ELISA Development

  • Published By:
  • Category: Health & Fitness
  • Published Date: February 10, 2021
  • Modified Date: February 10, 2021
  • Reading Time: 4 Minutes

Featured Image Caption: ELISA Laboratory Procedures

For biological assays, having them cultured correctly is all there is to have them function properly. And with ELISA, you can stay ahead of everything regarding biological samples. Many issues can arise when working with biological samples, and it could be worse if the format isn’t defined, which is the main aim of ELISA. To help with analyzing, detecting, and monitoring the progress of immunological assays.

Need help with understanding everything that has to do with this aspect of biology? You can check out the ELISA troubleshooting guide for assay development in laboratory procedures.

What is ELISA?

Short for enzyme-linked immunosorbent assay, ELISA is an immunological assay that is prepared in the lab to help measure glycoproteins, proteins, antigens, and antibodies. It is used widely in medical practice, especially when it comes to laboratory tests that involve detailed scrutiny.

An example of such a scenario is in pregnancy tests, HIV infection, and measurement of soluble receptors in serum.

The ideal procedure involves the use of 96 immune plates, which allows you to test multiple samples at once. A select type of plate is required to record success with the procedure. You want the antigen or antibody to stick to the surface of the plate, and this is only possible if you are using the right kit.

Common Problems with ELISA Assays

common elisa problems

Common ELISA Problems

For the most part of it, most biological procedures can take place if the right conditions are met, and for ELISA, there are technical issues that could put the process at risk. Once the samples have been collected and the captured antigen (control reagent) is placed on the plate’s surface, the antigen in the sample will bind to that of the control reagent. And in a situation where this doesn’t take place, you can expect to encounter some challenges with the procedure.

Some of the common problems with ELISA assays include

Picking the Right Antibody/Antigen

The antigen or antibody system needs to be carefully selected. It is crucial to choose the specific type that will bind easily with the sample. Two things are essential when picking the right antibody or antigen; you want to consider the sensitivity and quality of the specimen when making your choice.

Non-specific Binding Problem

Another possible problem you may encounter when working with biological assays is that of non-specific binding of the specimen to the surface. This is worse for systems that require complex samples as it is possible you will have to use dilatants, blockers, and protein stabilizers to help improve binding in the system.

Issue of False Positives

It could be dangerous to provide the wrong laboratory analysis of a test specimen, which is why the issue of false positives is a serious matter when dealing with ELISA assays. To solve this problem in immunoassays, you want to reduce cross-reactivity, which can be done with the help of stabilizers and diluting agents.

Increasing Assay Sensitivity and Specificity

Staying on top of the true and false positive outcomes of the procedure is an essential aspect of getting the job done correctly. And to do this, you want to ensure that the sensitivity and specificity are in their ideal state. You can make use of protein stabilizers to increase the levels in the system and, by so doing, improve the diagnostic accuracy of the procedure.

Keeping the Assay Stable

One problem you’ll also encounter in the lab is keeping the assay stable. This could be difficult if you have to keep the assay for a long time. You can find substrates and reagents that are useful in maintaining assay levels and keeping them safe on the shelf.

Given the best scenario, you want to ensure that other laboratory conditions are kept stable.

This includes the ideal temperature and lighting for keeping the samples steady. Most possibly, you want to isolate the reagents as far away from the culture materials. You also want to observe safety procedures such as proper handling and using the appropriate protective gear. There is more gist here https://www.thoughtco.com/important-lab-safety-rules-608156 on the essential rules to maintain in the lab.


ELISA assay could provide a detailed insight into immunological samples, and this has proven to be useful in laboratory technological. And for biological expert, knowing just how to avoid any complications in the course of lab analysis is paramount in ensuring the safety and accuracy of the procedure.

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