There are many protocols with a lot of adjuvants such as aluminium salts, squalene, freunds, etc. Can you give me your experience with how to choose the most comfortable adjuvant for total and separate (IgG, IgM, IgA) antibodies?
We have been using the type I (cholera toxin, E. coli LT) and the type II (LT-IIa, LT-IIb, LT-IIc) as strong mucosal and systemic (intradermal, peritoneal) adjuvants. These molecules work exceptionally well for augmenting immune responses to poor antigens.
Colleagues in Belfast (AFBI and Queen’s University) investigated Gerbu LQ 3000 and Montanide ISA 50V as replacements for Freund’s adjuvant for producing comparable polyclonals to small molecules (veterinary drugs) in rabbits. They found that Montanide produced no adverse effects in the animals and the related antibodies were of adequate sensitivity when compared to those from Freund’s rabbits. Montanide could be considered as a suitable replacement to Freund’s in this instance. The rabbits didn’t like Gerbu and antibody titres were low although sensitivities were high.
Comparison of three adjuvants used to produce polyclonal antibodies to veterinary drugs
Terence L. Fodey, Philippe Delahaut, Caroline Charlier, Christopher T. Elliott. Veterinary Immunology and Immunopathology 122 (2008) 25–34
Abstract
Two commercially available adjuvants, Gerbu LQ 3000 and Montanide ISA 50V, were assessed as potential replacements for Freund’s adjuvant by evaluating their efficacy in the production of polyclonal antibodies to veterinary drugs in rabbits. The aim was to find an adjuvant that could produce a similar (or enhanced) immune response in the host animal without the undesirable side effects associated with Freund’s complete and incomplete adjuvant. The assessment involved the examination of each injection site and the characterisation of the resultant antibodies with regards to antibody titre and sensitivity. It was found that the rabbits immunised with Gerbu adjuvant produced some of the most sensitive antibodies. However, titres were relatively low and adverse effects at injection sites were relatively common. Montanide adjuvant produced no adverse effects and the related antibodies were found to be of adequate sensitivity when compared to those from rabbits immunised with Freund’s. It was concluded that Montanide ISA 50V could be considered as a suitable replacement to Freund’s for the production of polyclonal antibodies, to low molecular weight compounds in rabbits.
You might try BLUE CARRIER® Hemocyanin from Concholepas concholepas (Biosonda). It has worked great for a variety of animals and I know it works very good in rabbits at least.
Dear Terry: I am very thankfull for your answer. However, I want to study about using bacteria as vaccine vehicle. So, I use LTB as target antigene.
Dear Kevin and Karen: In our laboratory, the cost for using special adjuvant is very limited. So, I have to consider about price of adjuvants which you recomment.
Whatever, thank for your answer.
Though many adjuvants are available these days, we are still using Freund's complete and incomplete adjuvant for polyclonals in rabbits. We therefore highly recommend the use of Freund's adjuvant for high titre antibodies in rabbits.
The adjuvant that has always worked in mice and in rabbits in our laboratory is Freund's complete and incomplete adjuvant.
I recommend you use freunds adjuvant including Complete and incomplete freunds adjuvant for both animals. 1st injection use Complete freunds adjuvant and 2-3 use incomplete freunds adjuvant. For the Ig type, you should understand the stage of each type which one produced first and after 2nd injection which one will produce or higher. My Ph.D thesis is mAb production.
cheer!!
I use titermax gold it works well but gives less side effects than Freunds
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