Lipopeptide adjuvants: generation of lactate dehydrogenase isoenzyme-specific antibodies for immunochemical diagnosis.
Lactate dehydrogenase catalyzes the final step in glycolysis, the interconversion of pyruvate and lactate. The tetrameric enzyme is composed of one or two subunits (H and/or M) resulting in five isoenzyme forms: LDH-H4, -H3M1, -H2M2, -H1M3, and -M4. The relative distribution of the LDH isoenzymes is tissue dependent and a significant marker for the diagnosis of hepatoma of the liver, myocardial infarction, muscular dystrophy, and a wide variety of other acute and chronic diseases to be detected by alterations of the LDH isoenzyme pattern in serum. Immunochemical approaches to the routine determination of LDH depend on isoenzyme specific antibodies. Since the H- and M-subunits for human LDH are highly homologous, LDH isoenzyme specific antibodies for immunochemical monitoring are hard to generate. Here we present data on the generation and characterization of LDH isoenzyme-specific mono- and polyclonal antibodies in different species in the presence of lipopeptide adjuvants. Western-Blot and ELISA analysis showed that antisera and monoclonal antibodies recognize their homologous antigens with high specificity and are therefore suitable for immunochemical monitoring of the LDH isoenzymes H4 and M4. In addition, they can be used for the determination of LDH isoenzyme specific activity which is an essential prerequisite for online amperometric immunosensor monitoring.
Gampp
T M
TM
Institut für Mikrosystemtechnik der Universität, AG Bioanalytische Mikrosysteme, Freiburg, Germany.
Moser
I
I
Jobst
G
G
Urban
G
G
Ayoub
M
M
Pfannes
S D
SD
Hoffmann
P
P
Bessler
W G
WG
Mittenbühler
K
K
D000276:Adjuvants, Immunologic; D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000378:metabolism; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D015151:Immunoblotting; D007527:Isoenzymes / Q000276:immunology / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000276:immunology* / Q000378:metabolism; D008074:Lipoproteins / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits
D000276:Adjuvants, Immunologic; D000906:Antibodies; D007527:Isoenzymes; D008074:Lipoproteins; D007770:L-Lactate Dehydrogenase