Development of monoclonal antibodies and simplified sandwich ELISA for canine neutrophil gelatinase-associated lipocalin A detection

2020 ◽  
Author(s):  
Jun Cao ◽  
Bo Tang ◽  
Xinyu Zhang ◽  
Xiaoli Xia ◽  
Huaichang Sun
Keyword(s):  
Monoclonal Antibodies ◽  
Sandwich Elisa ◽  
Kidney Injury ◽  
The Novel ◽  
E Coli ◽  
Neutrophil Gelatinase Associated Lipocalin ◽  
Hybridoma Cell Lines ◽  
Neutrophil Gelatinase ◽  
Canine Diseases ◽  
Good Agreement

Abstract Background Acute kidney injury (AKI) is one of the most common canine diseases with a high mortality rate. Neutrophil gelatinase-associated lipocalin (NGAL) is the novel biomarker for early diagnosis of renal injury. Only few sandwich ELISA kits are commercially available, all of which require the use of expensive enzyme-conjugated secondary or biotinylated antibody. The aims of this study were to develop high affinity monoclonal antibodies (mAbs) and simplified sandwich ELISA for canine NGAL detection. Results Recombinant canine NGAL was expressed in E. coli and purified to a high purity. Six hybridoma cell lines were generated by immunization of mice with the purified protein, all of which secreted high titers of specific mAbs. By screening 36 different antibody combinations, a pair of mAbs with high additivity and P/N ratio was selected as the capture and detection antibodies. By conjugation of the detection mAb with horse radish peroxidase, a simplified sandwich ELISA was developed with a correlation coefficient of 0.9939, detection limit of 8.28 ng/mL. The parallel test of 42 samples from healthy and AKI dogs showed the good agreement in NGAL concentrations detected by the simplified sandwich ELISA and commercial ELISA kit. Conclusions We developed canine NGAL-specific monoclonal antibodies and simplified sandwich ELISA. The simplified sandwich ELISA could replace the commercial ELISA kits for canine NGAL detection with the advantages of reduced cost and detection time.

Neutrophil Gelatinase-associated Lipocalin as a Cardiovascular Biomarker

2011 ◽  
Vol 7 (3) ◽  
pp. 154
Author(s):  
Andrew K Roy ◽  
John Keaney ◽  
Niall G Mahon ◽  
Patrick T Murray ◽  
Catherine Macauley ◽  
...  
Keyword(s):  
Cardiovascular System ◽  
Kidney Injury ◽  
Aortic Aneurysms ◽  
The Novel ◽  
Neutrophil Gelatinase Associated Lipocalin ◽  
Endothelial Apoptosis ◽  
Abdominal Aortic ◽  
Research Findings ◽  
Cardiovascular Biomarker ◽  
Neutrophil Gelatinase

Neutrophil gelatinase-associated lipocalin (NGAL) is a 25 kDa glycoprotein found in many tissues, and has attracted increasing attention as a biomarker for the early diagnosis of acute kidney injury (AKI). In the kidney, it is produced and released in response to ischaemic insults, and thus may also play a role in diagnostic and prognostic evaluation and management of patients with AKI, including those with acute cardiorenal syndromes. In the cardiovascular system, NGAL has been implicated in endothelial apoptosis and atherosclerosis, as well as in the thrombi associated with abdominal aortic aneurysms. This review explores the novel roles of NGAL in the cardiovascular system, and highlights some of the key research findings in this rapidly evolving field.

Neutrophil Gelatinase-associated Lipocalin as a Cardiovascular Biomarker

2011 ◽  
Vol 6 (2) ◽  
pp. 118
Author(s):  
Andrew K Roy ◽  
John Keaney ◽  
Niall G Mahon ◽  
Patrick T Murray ◽  
Catherine Macauley ◽  
...  
Keyword(s):  
Cardiovascular System ◽  
Kidney Injury ◽  
Aortic Aneurysms ◽  
The Novel ◽  
Neutrophil Gelatinase Associated Lipocalin ◽  
Endothelial Apoptosis ◽  
Abdominal Aortic ◽  
Research Findings ◽  
Cardiovascular Biomarker ◽  
Neutrophil Gelatinase

Neutrophil gelatinase-associated lipocalin (NGAL) is a 25 kDa glycoprotein found in many tissues, and has attracted increasing attention as a biomarker for the early diagnosis of acute kidney injury (AKI). In the kidney, it is produced and released in response to ischaemic insults, and thus may also play a role in diagnostic and prognostic evaluation and management of patients with AKI, including those with acute cardiorenal syndromes. In the cardiovascular system, NGAL has been implicated in endothelial apoptosis and atherosclerosis, as well as in the thrombi associated with abdominal aortic aneurysms. This review explores the novel roles of NGAL in the cardiovascular system, and highlights some of the key research findings in this rapidly evolving field.

scholarly journals Clinical Application Neutrophil Gelatinase-Associated Lipocalin and Kidney Injury Molecule-1 as Indicators of Inflammation Persistence and Acute Kidney Injury in Children with Urinary Tract Infection

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Stanislava Petrovic ◽  
Natasa Bogavac-Stanojevic ◽  
Amira Peco-Antic ◽  
Ivana Ivanisevic ◽  
Jelena Kotur-Stevuljevic ◽  
...  
Keyword(s):  
Acute Kidney Injury ◽  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Kidney Injury ◽  
The Novel ◽  
Neutrophil Gelatinase Associated Lipocalin ◽  
Significant Difference ◽  
Kidney Injury Molecule 1 ◽  
Neutrophil Gelatinase

Background. The aim of this study was to examine the novel renal biomarkers neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) to assist pediatricians in the assessment of longer duration of inflammation and acute kidney injury (AKI) development during urinary tract infection (UTI).Methods. The patients enrolled in the study comprised 50 children (mean age was 6 months) with UTI. NGAL in serum and urine (sNGAL and uNGAL, resp.) and KIM-1 in urine were measured by enzyme-linked immunosorbent assays.Results. uNGAL levels in subjects with longer duration of inflammation were higher (115.37 ng/mL) than uNGAL levels in subjects with shorter duration of inflammation (67.87 ng/mL,P=0.022). Difference in sNGAL and KIM-1 levels was not significant (P=0.155andP=0.198, resp.). Significant difference was seen in KIM-1 excretion among groups with and without AKI (P=0.038). KIM-1 was not able to discriminate between subjects with and without AKI (area under the curves (AUC) = 0.620,P=0.175).Conclusions. uNGAL cannot be used for screening of the duration of inflammation during UTI. Accuracy of KIM-1 in screening of AKI development in children with UTI is low. We suggest larger studies to check the negative predictive value of KIM-1 for the development of AKI.

Evaluation of urinary neutrophil gelatinase-associated lipocalin and urinary kidney injury molecule-1 as biomarkers of renal function in cancer patients treated with cisplatin

2020 ◽  
Vol 26 (7) ◽  
pp. 1643-1649
Author(s):  
Elliyeh Ghadrdan ◽  
Sholeh Ebrahimpour ◽  
Sanambar Sadighi ◽  
Samira Chaibakhsh ◽  
Zahra Jahangard-Rafsanjani
Keyword(s):  
Acute Kidney Injury ◽  
Serum Creatinine ◽  
Characteristic Curve ◽  
Kidney Injury ◽  
Acute Kidney Injury Network ◽  
The Novel ◽  
Urinary Ngal ◽  
Neutrophil Gelatinase Associated Lipocalin ◽  
Kidney Injury Molecule 1 ◽  
Neutrophil Gelatinase

Introduction Cisplatin-associated acute kidney injury (AKI) is the major limitation to the use of cisplatin-based chemotherapy regimens. Serum creatinine as a traditional marker did not increase in a timely enough fashion in AKI patients. Therefore, recently, the novel markers such as neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) were considered for early detection of AKI. The aim of this study was to compare the sensitivity and specificity of urinary NGAL and KIM-1 with serum creatinine in cisplatin related AKI. Methods Patients ≥18 years with solid tumors who received cisplatin-based chemotherapy were included. Urine samples were collected 0, 6 and 24 h after cisplatin infusion and the urinary NGAL, KIM-1, and creatinine concentrations were evaluated. NGAL and KIM-1 concentrations were adjusted based on urine creatinine to eliminate hydration effects. Serum creatinine levels were assessed at the base and 72 h after cisplatin administration. Results Seven out of the 35 recruited patients (20%) suffered from AKI defined by Acute Kidney Injury Network criteria. In AKI patients, the ratio of urinary KIM-1–creatinine at 24 h compared to baseline (24 h/baseline) and NGAL–creatinine 24 h/baseline were significantly higher than those of non-AKI group ( p = 0.037 and 0.047 respectively). The area under the receiver-operating characteristic curve for KIM-1–creatinine 24 h/baseline and NGAL–creatinine 24 h/baseline were 0.78 (0.59–0.96, p = 0.032) and 0.77 (0.57–0.97, p = 0.036) respectively. Conclusions Our findings showed that the changes in urinary NGAL–creatinine and KIM-1–creatinine ratios, 24 h after cisplatin administration can be utilized to predict AKI in cisplatin recipients.

Application of Monoclonal Antibodies to Dulcitol 1-Phosphate Dehydrogenase for Rapid Detection of Salmonella

1995 ◽  
Vol 58 (8) ◽  
pp. 847-852 ◽  
Author(s):  
TAKAHISA MIYAMOTO ◽  
HUAIZE TIAN ◽  
KIYOSHI MATSUNO ◽  
RYOJI TAKATA ◽  
SHOJI HATANO
Keyword(s):  
Monoclonal Antibodies ◽  
Salmonella Typhimurium ◽  
Magnesium Chloride ◽  
Rapid Detection ◽  
Limit Of Detection ◽  
Sandwich Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
The Novel ◽  
Inoculum Level ◽  
E Coli

Monoclonal antibodies raised against dulcitol 1-phosphate dehydrogenase of Salmonella typhimurium IFO 12529 were screened against 20 serotypes of Salmonella and 13 non-Salmonella bacteria. A sandwich-capture, enzyme-linked immunosorbent assay (sandwich ELISA) was developed for detection of Salmonella in food. The assay utilizes two monoclonal antibodies (DUI2 and DU28) which show no cross-reactions with non-Salmonella bacteria. The limit of detection of the sandwich ELISA was about 1 × 107 CPU/ml. After cultivation in a medium containing dulcitol at 37°C for 18 h followed by the sandwich ELISA. 1 CPU of Salmonella was detected. Although a high inoculum level of E. coli interfered with the detection of Salmonella, the interference was minimized by using a selective dulcitol-magnesium chloride-pyridinesulfonic acid medium for enrichment. The novel ELISA procedure detected Salmonella in chicken filtrates inoculated with 1.4 CPU/50 m1 and 1.3 × 107 CPU/50 ml of E. coli within 25 h.

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