antibody labelling for immunochromatographic assay - (Nov/08/2007 )
Hi
Does anyone know a good procedure to label antibody fragment with gold colloid for lateral flow assay? I don't have any problem labelling monoclonal antibodies, but I can't seem to be able to label scFv or Fab fragment. I know that for optimum labelling the pH of the gold solution should be about 1 above the pI of the antibody fragment. However, it seems impossible to change the pH of the gold colloid solution.
Any advives would be greatly appreciated, thanks a lot
Marie
-mlb-
QUOTE (mlb @ Nov 8 2007, 08:50 AM)
Hi
Does anyone know a good procedure to label antibody fragment with gold colloid for lateral flow assay? I don't have any problem labelling monoclonal antibodies, but I can't seem to be able to label scFv or Fab fragment. I know that for optimum labelling the pH of the gold solution should be about 1 above the pI of the antibody fragment. However, it seems impossible to change the pH of the gold colloid solution.
Any advives would be greatly appreciated, thanks a lot
Marie
Does anyone know a good procedure to label antibody fragment with gold colloid for lateral flow assay? I don't have any problem labelling monoclonal antibodies, but I can't seem to be able to label scFv or Fab fragment. I know that for optimum labelling the pH of the gold solution should be about 1 above the pI of the antibody fragment. However, it seems impossible to change the pH of the gold colloid solution.
Any advives would be greatly appreciated, thanks a lot
Marie
You should adjust the pH of protein solution and do a titration using different protein to gold particles. Anal Biochem has a paper on how to do this.
-genehunter-1-
QUOTE (genehunter-1 @ Nov 8 2007, 05:18 PM)
QUOTE (mlb @ Nov 8 2007, 08:50 AM)
Hi
Does anyone know a good procedure to label antibody fragment with gold colloid for lateral flow assay? I don't have any problem labelling monoclonal antibodies, but I can't seem to be able to label scFv or Fab fragment. I know that for optimum labelling the pH of the gold solution should be about 1 above the pI of the antibody fragment. However, it seems impossible to change the pH of the gold colloid solution.
Any advives would be greatly appreciated, thanks a lot
Marie
Does anyone know a good procedure to label antibody fragment with gold colloid for lateral flow assay? I don't have any problem labelling monoclonal antibodies, but I can't seem to be able to label scFv or Fab fragment. I know that for optimum labelling the pH of the gold solution should be about 1 above the pI of the antibody fragment. However, it seems impossible to change the pH of the gold colloid solution.
Any advives would be greatly appreciated, thanks a lot
Marie
You should adjust the pH of protein solution and do a titration using different protein to gold particles. Anal Biochem has a paper on how to do this.
thanks for the info, would you have any description of the article? I have been looking for it but can't seem to find it.
Many thanks
-mlb-
Liu et al Anal Biochem 147, 49-56, 1985 and ref within. good luck.
-genehunter-1-
QUOTE (genehunter-1 @ Nov 8 2007, 06:12 PM)
Liu et al Anal Biochem 147, 49-56, 1985 and ref within. good luck.
Thank you so much for your help
-mlb-