#Hormonology
Section: Theory
Subject: Eliza
1. One of the methods of identifying antigen or antibody is ELISA, which is used in both serology and hormonology (hormone measurement).
2. It is the same as the biochemistry test, but there is one difference. In your biochemistry, you have a test tube (to which we add serum) and a standard tube (to which we add a calibrator solution, which we know the concentration of), and with the help of a spectrophotometer, we calculate and find the concentration of the unknown substance in the serum through the OD reading. we do
But aaaaaaaaaaaaaaaaaaaaaaaaaaa
In ELISA, a test + several standards with different and specific concentrations + a cut-off (control)
And with the help of Eliza Riederba, he gives us the specific wavelength of both OD and concentration
The case is related to Ag_Ab complex. And we use enzyme for this test, that is
The precursor is converted into a colored product by the enzyme, and we find its OD and concentration with ELISA reader
3. Types of Eliza:
Before that, they add the contents in a series of very small containers called wells (wells or microplates), but the issue is that depending on the type of ELISA, the inner part of the bottom of the well is industrially coated with antigen or antibody. For example, if the antigen is coated, we should look for antibodies in the patient’s serum. If the antibody is coated, we look for the antigen in the serum.
Direct ELISA:
It has two modes:
1_ The bottom of the well is coated with antigen:
Procedure: Serum (provided it has Ab) is produced with serum antibody that binds with the labeled enzyme and the product is read.
2_ The bottom of the well is coated with antibody: on the contrary.
This ELISA model is not used much, it is more of a research aspect
Indirect ELISA:
The bottom of the well is coated with antigen, then it is diluted with serum (provided that you have Ab, because the serum is diluted and contains antibodies, so called antibody titer)
clarification:
Here, the antigen binds with the serum antibody
This is the primary antibody. be careful
Now, the enzyme is conjugated to the secondary antibody (human globulin antibody) and binds with the complex (it is attached to the Fc part of the primary antibody) to produce a colored product and check
As we talked about the antibody titer, this method is used in serology and diseases and helps to measure Ig.
ELISA Sandwich:
The antibody is coated on the bottom of the well, so it binds with the serum antigen
clarification:
Primary antibody + antigen + enzyme conjugated with secondary antibody, antigen sandwiched between two antibodies, so-called
This method is used in hormonology
For example: HCG, PSA, LH, FSH, TSH
Competitive ELISA (Competition ELISA):
There are two modes, for example, the bottom of the antigen well is coated
Antigen + primary antibody + enzyme conjugate
Competition between primary and secondary antibody, so here is competition
If Ab is primary > Ab is secondary, it is the same
If the primary Ab
