Detection of Cystatin C biomarker for clinical measurement of renal disease by developed ELISA diagnostic kits

Abstract

BackgroundHuman cystatin C (HCC) is a potential biomarker for tubular damage and impaired renal function. It is difficult to obtain efficient paired monoclonal antibodies against HCC with low molecular to meet the requirements for clinical application The present study was to establish a stable and repeatable measurement for HCC with self-made monoclonal antibodies (McAbs) and Variable domain of heavy chain of heavy-chain antibody (VHHs) increase the sensitivity.MethodsWith hybridoma technology and phage display technology: R-HCC as a screening antigen and N-HCC as the detector for antigens to obtain the specific antibody and established an enzyme-linked immunosorbent assay for human cystatin C using self-made McAbs and VHHs.ResultsWe have successfully obtained three McAbs; 5 F2, 4E4, 1E11 and four VHHs; 3-2, 3-24, 3-33 and 4-5 which were specific for HCC. The measurement of HCC was established with the self-made monoclonal antibodies and VHHs with a high sensitivity the lower limit of detection at 0.5 ng/ml and the detection range at 0.5 ~ 31.3 ng/ml.ConclusionOur data provides a new approach for paired antibody screening and testing of the small molecular biomarker with a single dominant epitope, with the important biological and clinical significance.

Glycosylation is important for legumain localization and processing to active forms but not for cystatin E/M inhibitory functions

“…In recent years, ELISA has been considered as the standard quantitative method for measurement of several proteins in diseases, such as cytokines [178], cathepsin L and K [179,180], and serum levels of cystatin C have clinical value in estimation of glomerulus filtration rate [181,182]. A commercially available human legumain ELISA-duokit (DY4769) from R&D Systems, containing a biotinylated legumain pAb for detection, was used to measure the total legumain concentrations in plasma or serum obtained from patients with carotid atherosclerosis, STEMI and CVD patients, healthy controls, and in conditioned media from primary cells and cell lines (paper I-III).…”

Establishment of Novel Urinary Kidney Disease New Biomarkers and Therapeutic Effect of Methanol Fraction of Terminalia Arjuna on Acetaminophen Induced Kidney Disease in Rats

“…SOD, catalase, MDA level of kidney tissue measured in UV-VIS Spectrophotometer by standard protocol [9,15,16]. IL-18 [17,18], KIM-1 [18,19], and cystatin-C [20] were measured by commercially available standard kit with standard protocol for kit literature by an ELISA plate reader.…”

Salivary proteome profile of women during fertile phase of menstrual cycle as characterized by mass spectrometry

“…Likewise, Cystatin-A and cystatin isoforms were reported in gingival crevicular fluid in periodontal patients and chicken egg white during embryogenesis, respectively [50,51]. It is suggested that salivary glands respond to inflammation stimuli to secrete more Cystatin-C into saliva [50] and it has been considered as a biomarker candidate of renal function [52]. Alterations of serum Cystatin C was considered as early marker for hyperthyroidism, cancer, renal function in diabetic patients, and cardiovascular diseases [53][54][55][56].…”