Dear Folks,
We immunized mice with 7kd synthetic peptides. We could not get a very good anti-body titer in first booster dose (second time immunization). However, we got satisfactory antibody response in second booster dose (third time immunization). Why the anti-body titer raises in second booster dose? we expected better antibody response in first booster dose itself.
Is it OK to get low antibody titer after first booster dose?
Any explanation for this.
Hello Vijayamahentesh
There are several issues related to the peptides. When the peptide is injected into mice it is not necessary that this injected peptide will follow the same route of processing as any other pathogen, and therefore may not give the same immune response as you would usually expect. This is because the peptide is smaller, linear and more importantly it is non-conformational. For the injected peptide to activate B cells to produce antibody response it needs to possess a conformation by incorporating conformational elements.
Thank you.
Thank you Malcolm for your answer.
What are conformational elements and how to incorporate them.
Did you inject the peptide only or have you conjugated it to a carrier protein (e.g. KLH) and have you used adjuvants? Which?
No conjugation is used.. direct peptide with CFA in priming and IFA in the subsequent booster dose.
Hello Vijayamahantesh
I agree with Wolfgang Schechinger. In your case you should go for conjugating your peptide with carrier protein to get a higher immune response. The routinely used carrier proteins are keyhole limpet haemocyanin (KLH), bovine serum albumin (BSA). There are other carrier proteins which you could use like ovalbumin and thyroglobulin.
Conformation is basically the shape of the molecule. It is a tedious and time consuming process which involves protein chemistry.
Thank you.
Thank you Malcolm.
I will repeat the experiment with carrier protein.
Dear Vijayamahantesh, I fully agree with the suggestions, that a carrier protein is important. In addition I like to mention, that the formation of immunological memory needs time and is often depending on the priming antigen dose. A high antigen dose for priming is usually not optimal for memory formation.
Willem B. Van Muiswinkel ,
I recently joined the lab. My mentor has this problem. They outsourced the generation of antibody. The antibody producing company gave primary dose on DAY 0 and the booster dose on DAY 14. I feel they have given booster dose too early. What is your thought on this. @
Dear Vijayamahantesh, It is very well possible, that a booster dose after 14 days is too early. In the added file you will see a study by Benner et al (1974). They observed an effect of the priming antigen dose and an optimal interval of 2-3 months between first and second antigen injection in mice. Hopefully this will be helpful in your study. All the best, Willem
Willem B. Van Muiswinkel
Thank you sir. It is very much useful for me.
- Badges
- Science topic
- Similar topics
- Clinical Pharmacology
- Dosing