RE: monoclonal antibody production

Woo, Gary (gary@amgen.com)
Tue, 1 Apr 1997 18:07:40 +0100

we would shoot up mice with a cell line that surface expressed the
protein we wanted an Ab against. it's kinda hit an miss. you can get
non-specific Abs or Abs against Ag you don't want. we used
microcytotoxicity or ELISA to screen for the Abs we wanted. Some
proteins are very antigenic and you get Abs on the first try. Other Ags
produce less of a response and can take quite a few immunizations to
produce a good, specific Ab. if you want any specifics or references on
our procedure drop me an email

hope this helps
gary woo

>----------
>From: Meenakshi Roy
>Sent: Tuesday, April 1, 1997 5:30
>To: Cytometry Mailing List
>Subject: monoclonal antibody production
>
>
>Dear flow-ers,
> This is not a directly flow related question. I am planning to produce
>monoclonal antibodies to certain cloned proteins.
> My question is, what form of a protein is most effective for monoclonal
>antibody production? I have been told that fusion proteins dont always
>fold similarly to the native protein. So, monoclonal antibodies that
>recognise the fusion protein, dont always recognise the native protein. On
>the other hand, immunisation with transfectants can result in many
>antibodies that are non-specific to the protein of interest.
> Also, what are the relative merits of HA tagged fusionproetins, or Ig
>chimera fusion proteins, or c-myc tagged fusion protiens for this purpose?
> Any experiences, suggestions shall be highly appreciated,
> Thanks in advance,
>Dr. Meenakshi Roy
>Stanford university
>
>
>