Reiter Protein Antigen: I. RELATIONSHIP OF SOME PHYSICAL AND CHEMICAL CHARACTERISTICS TO SEROLOGICAL ACTIVITY

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Abstract
The serologically active component of Reiter protein antigen considered by D''Alessandro and his co-workers to be a soluble protein was found to be particulate, with particle sizes ranging from 100 to 800 m[mu] in a given lot of antigen. Different lots which had the same serological titer contained variable amounts of non-reactive protein and carbohydrate. Since it is desirable to have an antigen in the greatest possible state of purity, purification was attempted by use of controlled pH and ionic strength buffers in an effort to selectively precipitate or separate the reactive and non-reactive components. The reactive protein was recovered from the precipitate when an ionic strength of 0.01 and pH values of 4 and 5 were employed. The reactive protein could also be adsorbed to diethylaminoethyl cellulose at pH 7, and eluted by use of buffers at pH 5 and ionic strength 1. Differential migration with isopycnic gradient centrifugation employing sucrose demonstrated that the serologically reactive component had a density of approximately 1.4. The reactive components and part of the non-reactive components could be sedimented by centrifugation at approximately 30,000 R.C.F.