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A monoclonal antibody (mAb), ASPM-2, produced against N-(gamma-maleimidobutyryloxy)-succinimide (GMBS)-conjugated polyamine spermine [Spm; Fujiwara et al. (1994) Histochemistry 102, 397-404] was used for the development of an enzyme-linked immunosorbent assay (ELISA) for acetylpolyamines (Ac-PAs) in human urine. The ELISA is based on the principle of competition between an analyte and Spm-glutaraldehyde-bovine serum albumin conjugate-coated polystyrene microtiter wells for the mAb, followed by immunoreaction with biotinylated anti-mouse immunoglobulin and horseradish peroxidase-streptavidin. The ASPM-2 mAb showed strong immunoreaction with N1,N12-diacetylspermine (2Ac-Spm), N-monoacetylspermine (Ac-Spm), and N1-acetylspermidine (N1-Ac-Spd), the EC50 values being 29, 50, and 51 microM, respectively, but no cross-reaction with other PA-related compounds or amino acids. The method was used to measure urinary Ac-PA levels in healthy subjects and cancer patients, without pretreatment of the specimens, mean concentrations of 3.25 and 2.80 mumol per 24-h urine, respectively (as N1-Ac-Spd), being found. The ASPM-2 ELISA for N1-Ac-Spd, which is the PA most relevant to the analysis of human urine among the three Ac-PAs mentioned above, is specific and accurate, and can easily be used to analyze large numbers of specimens in parallel. It should thus have potential for studying the relationship between urinary N1-Ac-Spd levels and cancer.
K Fujiwara, and
H Kanetake, and
K Furukawa, and
Y Masuyama, and
G Bai, and
H Tanimori, and
T Kitagawa
September 2018,
Planta medica,
K Fujiwara, and
H Kanetake, and
K Furukawa, and
Y Masuyama, and
G Bai, and
H Tanimori, and
T Kitagawa
December 1988,
Journal of immunological methods,
K Fujiwara, and
H Kanetake, and
K Furukawa, and
Y Masuyama, and
G Bai, and
H Tanimori, and
T Kitagawa
September 1979,
Canadian journal of microbiology,
K Fujiwara, and
H Kanetake, and
K Furukawa, and
Y Masuyama, and
G Bai, and
H Tanimori, and
T Kitagawa
February 1990,
Clinical chemistry,
K Fujiwara, and
H Kanetake, and
K Furukawa, and
Y Masuyama, and
G Bai, and
H Tanimori, and
T Kitagawa
January 2024,
Methods in molecular biology (Clifton, N.J.),
K Fujiwara, and
H Kanetake, and
K Furukawa, and
Y Masuyama, and
G Bai, and
H Tanimori, and
T Kitagawa
September 1991,
Cell proliferation,
K Fujiwara, and
H Kanetake, and
K Furukawa, and
Y Masuyama, and
G Bai, and
H Tanimori, and
T Kitagawa
September 1988,
Journal of clinical microbiology,
K Fujiwara, and
H Kanetake, and
K Furukawa, and
Y Masuyama, and
G Bai, and
H Tanimori, and
T Kitagawa
November 2017,
Journal of fish diseases,
K Fujiwara, and
H Kanetake, and
K Furukawa, and
Y Masuyama, and
G Bai, and
H Tanimori, and
T Kitagawa
March 1981,
Immunitat und Infektion,
K Fujiwara, and
H Kanetake, and
K Furukawa, and
Y Masuyama, and
G Bai, and
H Tanimori, and
T Kitagawa
January 1997,
The Southeast Asian journal of tropical medicine and public health,
