Determination of a peptide in urine of patients after allogeneic hematopoietic stem cell transplantation by LIF-CE-IA
10.3760/cma.j.issn.1009-9158.2010.10.015
- VernacularTitle:毛细管电泳竞争免疫分析法测定异体干细胞移植患者尿液中小肽分子
- Author:
Zuojun SHEN
;
Xiaodong HE
;
Maoli YI
;
Zimin SUN
- Publication Type:Journal Article
- Keywords:
Stem cell transplantation;
Urinalysis;
Peptides;
Electrophoresis,capillary
- From:
Chinese Journal of Laboratory Medicine
2010;33(10):963-966
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop an assay with polyclonal antibodies against a fragment derived from human albumin for determination of a peptide in urine, and to provide an early diagnostic tool for GVHD. Methods A small peptide composed of 16 amino acids (LVRYTKKVPQVSTPTL) was synthesized artificially. The immunogen was then diluted into 100 μg/kg body weight of rabbit. Subcutaneous injection in the immune animals was performed on both sides of spine and groin with 2.5 ml antigen suspension for three times, in order to prepare the polyclonal antibodies. The peptide antigen was then labeled with fluorescein isothiocyanate (FITC), and detected by LIF-CE-IA with the pre-prepared antibodies. Results The titer of serum. The migration time of the labeled antigen was 5.93 min, while the migration time of antigen-antibody complex was 6.47 min. The linear range of the method was 16 to 512 ng/ml, and the minimum detection limit was 10 ng/ml. Conclusions The polyclonal antibodies against the peptide antigen was isolated successfully, which possessed high titer and specificity. These results indicated that the assay was simple and rapid and could be applied for the early diagnosis of patient with GVHD.
