Abstract: A method for quantification of a titer of a neutralizing antibody to a neurotoxin is disclosed. The method comprises the steps: (a) mixing a standard sample containing a fixed amount of a neurotoxin and a test sample containing a neutralizing antibody to said neurotoxin; (b) administering the mixture obtained in step (a) into the muscle of a non-human mammal; (c) applying electric stimulus to said non-human mammal; (d) measuring a compound muscle action potential (CMAP) due to contraction of the muscle of said mammal by application of electric stimulus with an electromyograph; and (e) analyzing CMAP amplitude data obtained in step (d) for an extent of decrease in amplitude by a non-neutralized neurotoxin to thereby quantify a titer of the neutralizing antibody contained in the test sample.

Abstract: Disclosed is a method for preparing pertussis toxin toxoid by a treatment pertussis toxin with formaldehyde, which comprises carrying out said treatment in the presence of lysine or glycine in combination with N-acetyltriptophan. The resultant toxoid does not exhibit toxicity reversion, thereby making the toxoid highly suitable for use in pertussis vaccines.

Abstract: Improved method for the purification of LPF-HA (Leucocytosis promoting far hemagglutinin) on industrial scale which comprises contacting a LPF-HA-containing solution from culture media of Bordetella pertussis with a cellulose sulfate gel, a crosslinked polysaccharide sulfate gel, or a polysaccharide gel chemically bound with dextran sulfate, thereby adsorbing LPF-HA on the gel, and then eluting LPF-HA from the gel. Said method can give a highly purified LPF-HA which does not contain any other proteins, lipid, saccharides, etc. and further undesirable endotoxin, and hence can be used for producing various reagents, medicines and pertussis vaccine.

Abstract: A method for production of HA fraction containing protective antigens of Bordetella pertussis (i.e. a fraction containing F-HA and LPF-HA) in an industrial scale, for instance, with a fermentater, which comprises inoculating a strain of B. pertussis in a liquid culture containing a cyclodextrin or its derivative, culturing it by a spinner culture under controlling the culture temperature and the amount of dissolved oxygen and under defoaming condition, optionally under controlling the pH range, and harvesting the produced HA fraction from the culture broth at a stage of from logarithmic growth phase to static grow phase, and a method for production of a pertussis vaccine from the HA fraction thus obtained by formalinizing the HA fraction in the presence of an amino acid.

Abstract: Improved method for the purification of filamentous hemagglutinin (F-HA) on industrial scale which comprises contacting a culture of a microorganism of the genus Bordetella with a cellulose sulfate gel, a polysaccharide gel chemically bound with dextran sulfate, or a crosslinked polysaccharide sulfate gel, thereby adsorbing F-HA on the gel, and then eluting F-HA from the gel. Said method can give a highly purified F-HA which does not contain any other proteins, lipid, saccharides, etc. and further undesirable endotoxin, and hence can be used for producing various reagents, medicines and pertussis vaccine.