Determination of substance P in human nasal lavage fluid.
The determination of substance P (SP) concentrations in human nasal lavages can be used to monitor physiological and certain pathophysiological processes in human airway mucosa. But, because of the low concentrations, immunoassays of high sensitivity are needed. Two approaches to improve the sensitivity of the radioimmunological determinations of SP are compared: increasing the sample volume and miniaturizing the assay design. The characterization of SP-like immunoreactivity (SP-LIR) in human nasal lavage was performed by investigating the immunological specificity of the antibody used in the radioimmunoassays and by reversed-phase high-performance liquid chromatography separation of the SP-LIR. SP concentrations in nasal lavages can be reliably measured by each of the two introduced RIA methods. Despite the lower detection limit of the miniaturized immunoassay (0.2 in comparison to 1.3 fmol/incubate) it is advisable to increase the sample volume in order to improve the sensitivity because of the higher precision of the determinations. SP-LIR was found in nasal lavage specimens in concentrations between 2 and 10 fmol/ml and consisted of authentic SP and, to a less extent, SP-sulfoxide.
Schultz
K D
KD
Department of Clinical Immunology and Asthma OPD, Virchow-Klinikum, Humbolt University Berlin, Germany.
Furkert
J
J
O'Connor
A
A
Böttcher
M
M
Schmidt
M
M
Baumgarten
C R
CR
Kunkel
G
G
D002851:Chromatography, High Pressure Liquid; D006801:Humans; D017817:Nasal Lavage Fluid / Q000737:chemistry*; D010446:Peptide Fragments / Q000032:analysis; D011863:Radioimmunoassay / Q000379:methods / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D013373:Substance P / Q000032:analysis*
D010446:Peptide Fragments; D013373:Substance P