Ectoparasite saliva proteins and apparatus to collect such proteins

A composition comprising at least one isolated flea saliva product substantially free of contaminating material, said composition being produced by a process comprising:

• (a) collecting flea saliva products on a collection means within a saliva collection apparatus containing fleas, said apparatus comprising:
• (i) a housing operatively connected to a chamber, said chamber having an ambient temperature warmer than said housing thereby forming a temperature differential between said housing and said chamber, said housing being capable of retaining fleas;
• and
• (ii) an interface between said housing and said chamber, said interface comprising ((a)) a means capable of collecting saliva products deposited by fleas retained in said apparatus and ((b)) a barrier means capable of substantially preventing contaminating material from contacting said collection means, wherein said temperature differential attracts fleas retained in said housing to attempt to feed through said barrier means and collection means and, thereby, deposit saliva products on said collection means;
• and
• (b) extracting said saliva products from said collection means to obtain said composition.

A composition comprising at least one isolated flea saliva product substantially free of contaminating material, said composition being produced by a process comprising:

• (a) collecting flea saliva products on a collection means within a saliva collection apparatus containing fleas said apparatus comprising:
• (i) a housing operatively connected to a chamber, said chamber having an ambient temperature warmer than said housing thereby forming a temperature differential between said housing and said chamber, said housing being capable of retaining fleas;
• and
• (ii) an interface between said housing and said chamber, said interface comprising ((a)) a means capable of collecting at least a portion of saliva products deposited by fleas retained in said apparatus, and ((b)) a barrier means capable of substantially preventing contaminating material from contacting said collection means, wherein said temperature differential attracts fleas retained in said housing to attempt to feed through said barrier means and collection means and, thereby, deposit saliva products on said collection means;
• and
• (b) recovering said flea saliva product-containing collection means.

A composition comprising at least one isolated flea saliva protein, wherein said flea saliva protein is as isolated from a protein peak as represented in FIG. 2, said peak being selected from the group consisting of peak E, peak F, peak G, peak H, peak I, peak J, peak K, peak L, peak M and peak N, wherein said peak is produced by the method comprising:

• (a) collecting flea saliva products on a collection means within a saliva collection apparatus containing fleas said apparatus comprising:
• (i) a housing operatively connected to a chamber, said chamber having an ambient temperature warmer than said housing thereby forming a temperature differential between said housing and said chamber, said housing being capable of retaining fleas;
• and
• (ii) an interface between said housing and said chamber, said interface comprising ((a)) a means capable of collecting at least a portion of saliva products deposited by fleas retained in said apparatus, and ((b)) a barrier means capable of substantially preventing contaminating material from contacting said collection means, wherein said temperature differential attracts fleas retained in said housing to attempt to feed through said barrier means and collection means and, thereby, deposit saliva products on said collection means;
• (b) extracting said products from said collection means to form said flea saliva extract, by contacting said collection means with a first extraction solution of about 50% acetonitrile and about 1.0% trifluoroacetic acid, collecting a flea saliva product-containing first extraction solution, precipitating said extracted flea saliva products from said first extraction solution, resuspending said precipitated flea saliva products in an about 12.8% acetonitrile solution and recovering flea saliva products soluble in said about 12.8% acetonitrile solution to form flea saliva extract FS-1;
• (c) fractionating said flea saliva extract FS-1 to obtain separated peak fractions of flea saliva proteins by high pressure liquid chromatography under conditions equivalent to use of a 15 cm.times.0.46 cm C4 column and a gradient of from about 0.1% trifluoroacetic acid in water to about 0.085% trifluoroacetic acid in 90% acetonitrile, at a flow rate of 0.8 milliliters per minute;
• and
• (d) recovering at least one of said peak fractions substantially free of the remaining fractions to obtain said peak.