Hi, I'm Kaniz, PhD student at university of Leeds. I'm working with HBc VLPs to generate different vaccine scaffolds which are expressed in E. Coli. I have found that the HBc VLPs contain a good amount of nucleic acids (NA) within the VLPs.
My questions are:
1. How much NA can be present in vaccine for safe use for human (and also for animal experiment)?
2. How can we determine whether the NA is DNA or RNA?
Please let me know if someone knows the answers of my questions.
Thank you
hi
I don't think you have a problem with this. you can run 20 ul of routine bacterial vaccine (such as cough -typhoid -cholera vaccine) sample with Ethidium bromide in 1% agarose gel and Compared to your own vaccine . Also, the presence of nucleic acids in the vaccine plays a similar role of CpG in stimulating the immune system.
good luck
I agree with Gerritzen, you can treat the vlps with approved nucleases like benzonase or micrococcal nuclease.
For more details regarding residual NAs in vaccines, kindly refer the following WHO guidelines.
https://www.who.int/biologicals/Molecular%20Methods%20Final%20Mtg%20Report%20April2005.pdf?ua=1
Best
Dear Gerritzen, thank you very much for the information.
After lysis of the cells, I always treat the soluble proteins with nuclease to reduce/remove DNA. As some of the VLPs (I'm working with) contain the nucleic acid binding domain, therefore, the VLPs contain moderate amount of DNA (measured by nanodrop).
Dear Raju, thank you for your answer. I agree with you.
Many many thanks for the link.
Regards,
Kaniz
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