Assays of urine levels of HNL/NGAL in patients undergoing cardiac surgery and the impact of antibody configuration on their clinical performances

Cai, Linjun; Borowiec, Jan; Xu, Shaoyuan; Wang, Han; Venge, Per

doi:10.1016/j.cca.2009.01.030

Cited by 52 publications

(57 citation statements)

References 33 publications

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“…Several research groups used one of these assays to determine the levels of HNL/NGAL in urine and drew the conclusion that HNL/NGAL is a biomarker of AKI (12,13,16 -18). Our previous results indicated that the antibody configuration had an effect on the clinical performance of the assay (19,25). We also reported, for the first time, the existence of several molecular forms of HNL/NGAL in urine obtained from patients after cardiac surgery and that the presence of dimeric and monomeric forms and their ratios changed after operation (19).…”

mentioning

confidence: 82%

“…In addition, we determined HNL/NGAL concentrations using our previously published monoclonal antibody (mAb)697-polyclonal antibody-based ELISA (19). The basic protocols for these five ELISAs are the same except for the specific antibodies (Diagnostics Development, Uppsala, Sweden) used in the assay.…”

Section: Sensitive Elisas For Hnl/ngal Quantificationmentioning

confidence: 99%

“…HK-2-conditioned supernatants at the 72-hour time point were harvested and supplemented with 0.1 mM phenylmethylsulfonyl fluoride (Sigma-Aldrich, Germany) and Complete protease inhibitor cocktail tablets (Roche, Germany). Nonreducing SDS-PAGE and Western blotting were performed according to the manufacturer's instructions and our previous paper (19).…”

Section: Western Blottingmentioning

confidence: 99%

“…Our previous results indicated that the antibody configuration had an effect on the clinical performance of the assay (19,25). We also reported, for the first time, the existence of several molecular forms of HNL/NGAL in urine obtained from patients after cardiac surgery and that the presence of dimeric and monomeric forms and their ratios changed after operation (19). The source of the different molecular forms of HNL/NGAL and what they might reflect has not yet been elucidated.…”

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confidence: 98%

“…Thus, the levels of HNL/NGAL were significantly increased in serum/plasma and urine after cardiac surgery and paralleled reduction in renal function (12,16,19). Several immunoassays have been developed for the measurement of HNL/NGAL.…”

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confidence: 99%

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The Origin of Multiple Molecular Forms in Urine of HNL/NGAL

Cai

Rubin

Wang

et al. 2010

Clinical Journal of the American Society of Nephrology

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Results: The major molecular form of HNL/NGAL secreted by neutrophils is dimeric, whereas the major form secreted by HK-2 cells is monomeric. This was reflected by a predominance of the monomeric form in urine from patients with AKI and the dimeric form in patients with UTIs. The epitope specificities of the antibody used in the ELISAs had a profound effect on assay performance and paralleled differences of the antibodies to identify the different forms of urine HNL/NGAL.Conclusions: The monomeric form is the predominant form secreted by tubular epithelial cells, and the dimeric form is the predominant form secreted by neutrophils. The development of molecular form-specific assays for HNL/NGAL may be a means to identify the origin of HNL/NGAL in urine and construct more specific tools for the diagnosis of AKI.

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mentioning

confidence: 82%

Section: Sensitive Elisas For Hnl/ngal Quantificationmentioning

confidence: 99%

Section: Western Blottingmentioning

confidence: 99%

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confidence: 98%

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confidence: 99%

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