Leishmania donovani:generation of monospecific antibody reagents to soluble acid phosphatase

Bates, P A and Kurtz, M K and Gottlieb, M and Dwyer, D M (1987) Leishmania donovani:generation of monospecific antibody reagents to soluble acid phosphatase. Experimental Parasitology, 64 (2). pp. 157-164. ISSN 0014-4894

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Abstract

Four monoclonal antibodies (McAbs) were generated against the soluble extracellular acid phosphatase (EC 3.1.3.2) (S-AcP) of Leishmania donovani. These were detected in the primary screen using an ELISA with promastigote culture supernatants as antigen. Three of the McAbs demonstrated bound S-AcP from such culture supernatants in an enzyme activity binding assay. All immunoprecipitated metabolically labeled S-AcP but none showed any binding to the promastigote surface by indirect immunofluorescence. Moreover, none reacted with Triton X-100 solubilized plasma membranes by immunoprecipitation or Western blotting. These results demonstrated that the McAbs did not recognize the surface membrane bound acid phosphatase, but were specific for the extracellular soluble enzyme. Further, none of the antibodies immunoprecipitated any of the five human acid phosphatase isozymes or reacted with them in Western blots or the enzyme activity binding assay. Therefore, they are specific for the parasite-derived enzyme. One of these was used to affinity purify sufficient L. donovani S-AcP to immunize a rabbit and generate a specific, polyvalent antiserum. This polyvalent antibody immunoprecipitated S-AcP activity but did not cross-react with the surface membrane acid phosphatase, indicating that these two parasite enzymes are separate gene products.

Item Type:
Journal Article
Journal or Publication Title:
Experimental Parasitology
Uncontrolled Keywords:
/dk/atira/pure/researchoutput/libraryofcongress/qr355
Subjects:
?? LEISHMANIA DONOVANIPROTOZOA, PARASITIC HEMOFLAGELLATE SOLUBLE ACID PHOSPHATASE (EC 3.1.3.2), (S-ACP)BIOMEDICAL AND LIFE SCIENCESIMMUNOLOGYPARASITOLOGYQR355 VIROLOGY ??
ID Code:
56071
Deposited By:
Deposited On:
24 Jul 2012 09:26
Refereed?:
Yes
Published?:
Published
Last Modified:
20 Sep 2023 00:22