How to determine neutralization amounts for peptide/antibodies?
To determine the amount of peptide needed to neutralize 1 ml of neat, undiluted antiserum: 1ml of antiserum contains 10mg of IgG. Among the 10mg total IgG, only 1% (under estimate) is against the specific antigen, therefore: (10mg)(1%)= 0.1mg or 100 mg of specific IgG against the antigen. Molecular weight of IgG=150,000 daltons, therefore, 100mg of IgG/150,000= 0.00066mmol or 0.66nmol 0.66nmol of antibody can neutralize twice the amount of peptide or 1.32nmol of specific peptide. Weight of peptide needed to neutralize = (1.32nmol)(M.W. of peptid)=X ng of peptide needed. To determine the amount of antibody needed to neutralize 1mg of peptide: 1mg of peptide/MW of peptide=X mmol (X times 1000nmol)1000 nmol of peptide needs 1000/2 nmol of specific IgG to be neutralized 1000/2 nmol of IgG= (1000/2 nmol)(MW of IgG)=Y ng of specific IgG. Since Y ng is the minimum needed, we recommend an overdose amount to be determined by end user.
