I have formulated antibody in semi solid dosage form and want to characterise its stability profile. Can someone help me how to quantify the amount of antibody in the formulation? how to get the antibody out of the formulation so it can be detected by ELISA or UV vis..
Dear Hanif, I don't know exactly what do you mean by semi solid dosage form, however if the problem is on how to quantify antibodies in a non purified sample, you could do it in an ELISA on anti-immunoglobulin antibodies, using dilutions of an isotype matching immunoglobulin of known concentration to create a reference curve.
If the problem is different, can you please give us some more detail?
Thank you Marcos
semi solid is a form of drug formulation that has similar characteristics to cream or gel products. I cant simply use this semi solid with ELISA, do you know how should i approach this to quantify the amount of my antibody in those formulation.
any references will be also good.
Thanks
So it seems the problem is to solubilise the semisolid formulation. What is the matrix made of? I guess it is not soluble in water: would it be possible to use some detergents, like NP-40 or Tween-20? Most antibodies can still bind up to 1% NP-40.
its made up of Polyethelene glycol, stearyl alcohol, cety alcohol, mineral oil, Xanthan gum and the antibody itself. what is the best method to extract the antibody from this compositions?
I don't have much experience with that kind of composition, but I would try to dilute it in some detergent. Buffers I use for immunoprecipitations are TNN (Tris-HCl pH 8 50mM, NaCl 250mM, NP-40 0.5%) or RIPA (Tris-HCl 50 mM pH 8, NaCl 150mM, NP-40 1%, NaDeoxicholate 0.5%, SDS 0.1%). In these buffers most antibodies still bind their antigens. If this works in solubilising the antibodies you can then try to quantify by ELISA on anti-immunoglobulin antibodies, as I suggested above.
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