I am working on a development project for ELISA using human sample pools and a newly developed antibody drug. In this assay we use one set of QCs containing ULOQ,HQC, MQC, LQC, LLOQ. And a sets of serially diluted Positive controls (STDs) with a similar pool and similar antibody drug.
I am having difficulty understanding the purpose of using QCs and how its related to our standard curve? What is the significance of that and based on the results ( recovery) what we can conclude and progress?
Id appreciate anyones insight.
As you have written that you are working on a development project for ELISA using human sample pools and a newly developed antibody drug. While doing so, its important to know and establish the various parameters like specificity, sensitivity, accuracy, stability etc for the developed ELISA.
The purpose of using QCs is to know that the developed ELISA is how much specific, sensitive and accurate to analyze the unknown samples (QCs). Normally, to establish that the ELISA developed by you must reach to the same concentration (that actually you do not know) of unknown QC sample provided for testing your ELISA set up.
Let me give you my example. As a team member of Prof. T.G. Shrivastav, former Head, Department of Reproductive Biomedicine, NIHFW, New Delhi,India), we developed several ELISA kits for protein and steroid hormones and we had this practice on routine basis. The quality control samples, we received from several international companies (for instance BIORAD) and run those QC samples (the concentrations were not revealed to us) in our ELISA. We sent the results of their QC samples. Thereafter, BIORAD analyzed our data (results) of their sample and gave us a gold certificate for our developed ELISA.
Let me give you little more clear example, BIORAD gave us several unknown samples (one of it was 11.20 ng/ml). We analyzed those QC samples in our ELISA and gave the result as (11.18 ng/ml). This data indicate that in terms of accuracy our developed ELISA meets almost >99% of BIORAD expectations.
This way one can evaluate the handwork and enjoy the successful development of ELISA kits with high accuracy. Indeed, several other parameters are also required to be checked in parallel in order to develop an ELISA kit for commercialization and patenting.
Best
Here is a nice summary of ELISA parameters which I hope can help unterstanding the principle:
Chapter Optimization, Validation and Standardization of ELISA
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