scholarly journals Development of two different formats of heterogeneous fluorescence immunoassay for bioanalysis of afatinib by employing fluorescence plate reader and KinExA 3200 immunosensor

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Ibrahim A. Darwish ◽  
Haitham AlRabiah ◽  
Mohammed A. Hamidaddin
Keyword(s):  
Effective Treatment ◽  
Solid Phase ◽  
Plasma Concentrations ◽  
Fluorescein Isothiocyanate ◽  
Small Cell ◽  
Clinical Settings ◽  
Fluorescence Immunoassay ◽  
Small Cell Lung ◽  
Plate Reader ◽  
Fluorescence Plate Reader

Abstract Afatinib (AFT) is a potent and highly selective drug used to treat various solid tumors including non-small cell lung cancer (NSCLC). To ensure safe and effective treatment of cancer patients with AFT, its plasma concentrations should be monitored. Thus, sensitive immunoassays are required for measuring AFT concentrations in plasma samples. In this study, two different formats of heterogeneous fluorescent immunoassays were developed and validated for AFT bioanalysis. These assays were microwell-based fluorescence immunoassay (FIA) using fluorescence plate reader and kinetic exclusion assay (KinExA) using KinExA 3200 immunosensor. Both FIA and KinExA were developed using the same reagents: mouse anti-AFT antibody, solid-phase immobilized AFT conjugated with bovine serum albumin protein (AFT-BSA), and goat anti-mouse IgG labelled with fluorescein isothiocyanate (FITC-IgG) for signal generation. The analytical performances of both assays were comparatively evaluated, and the results revealed that although both assays had comparable accuracies, KinExA was superior to FIA in terms of sensitivity and precisions. Moreover, both FIA and KinExA were better alternatives to the existing chromatographic methods for bioanalysis of AFT. The proposed FIA and KinExA are anticipated to effectively contribute in ensuring safe and effective treatment with AFT in clinical settings.

PD-L1 Gene Polymorphism and High Level of Plasma Soluble Pd-L1 Protein may be aSsociated with Non-Small Cell Lung Cancer

2015 ◽  
Vol 30 (4) ◽  
pp. 364-368 ◽  
Author(s):  
Sensen Cheng ◽  
Jinsong Zheng ◽  
Jingyan Zhu ◽  
Chao Xie ◽  
Xia Zhang ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Small Cell Lung Cancer ◽  
Cell Lung Cancer ◽  
Plasma Levels ◽  
Plasma Concentrations ◽  
Small Cell ◽  
Small Cell Lung ◽  
High Level ◽  
Nsclc Patients ◽  
L1 Protein

Background PD-1 and its ligand PD-L1 belong to the co-inhibition molecules, which can downregulate immune responses. The PD-L1 polymorphism and the level of soluble PD-L1 (sPD-L1) were investigated in non-small cell lung cancer (NSCLC). Methods A total of 288 NSCLC patients and 300 controls were enrolled. An A/C polymorphism at position 8923 in the PD-L1 gene was genotyped using the polymerase chain reaction-restriction fragment length polymorphism method. Results The prevalence of the 8923C allele was significantly higher in NSCLC patients than controls (10.2% versus 5.3%, p = 0.002, odds ratio 2.03, 95% confidence interval 1.30-3.17; data were adjusted for age and sex). NSCLC patients also showed increased plasma levels of sPD-L1 compared to controls (1.92 ng/mL versus 0.91 ng/mL, p<0.001). Furthermore, lung adenocarcinoma patients had higher sPD-L1 levels than patients with squamous cell carcinoma (p<0.01). However, no association was observed between the different genetic variants and plasma concentrations of sPD-L1. Conclusions The PD-L1 8923A/C polymorphism could be associated with increased susceptibility to NSCLC. Plasma levels of sPD-L1 are significantly increased in NSCLC patients, especially those with adenocarcinoma.

Elevated plasma concentrations of C-flanking gastrin-releasing peptide in small-cell lung cancer.

1989 ◽  
Vol 7 (12) ◽  
pp. 1831-1838 ◽  
Author(s):  
J J Holst ◽  
M Hansen ◽  
E Bork ◽  
T W Schwartz
Keyword(s):  
Cell Lines ◽  
Clinical Course ◽  
Plasma Concentrations ◽  
Molecular Size ◽  
Normal Subjects ◽  
Small Cell ◽  
Small Cell Lung ◽  
Extensive Disease ◽  
Lung Cancers ◽  
Gastrin Releasing Peptide

Many small-cell lung cancers (SCLCs) produce gastrin-releasing peptides (GRPs) (mammalian bombesin) but the plasma concentration of GRP is rarely elevated, possibly because of its rapid elimination. We developed a radioimmunoassay for the C-terminal flanking peptide of proGRP and measured its concentration in plasma from 71 patients with SCLC, in 27 healthy subjects and in 49 patients with other diseases including lung carcinomas. In addition, we studied the molecular size of immunoreactive C-flanking peptide in two SCLC cell lines and in plasma from SCLC patients. The concentration of immunoreactive C-flanking peptide in normal subjects and in control patients did not exceed 10 pmol/L and 26 pmol/L, whereas 72% of the SCLC patients had C-flanking peptide concentrations above 10 pmol/L. In patients with extensive disease (n = 35) the median concentration was 71 pmol/L (range, 10 to 940). ProGRP C-flanking peptide levels paralleled the clinical course in 12 patients. The molecule(s) responsible for the immunoreactivity had a molecular size of about 8 to 10 kd in both patient plasma and tumor cell lines, suggesting that the measured peptide(s) represented major fragment(s) if not the entire C-flanking peptide of proGRP. Thus this peptide(s) seems to be a useful marker for SCLC.

A polyclonal antiserum against chromogranin A and B - a new sensitive marker for neuroendocrine tumours

1990 ◽  
Vol 122 (2) ◽  
pp. 145-155 ◽  
Author(s):  
B. Eriksson ◽  
H. Arnberg ◽  
K. Öberg ◽  
U. Hellman ◽  
G. Lundqvist ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Chromogranin A ◽  
Neuroendocrine Tumours ◽  
Plasma Concentrations ◽  
Polyclonal Antiserum ◽  
Small Cell ◽  
Neuroendocrine Cells ◽  
Small Cell Lung ◽  
Pancreatic Tumours ◽  
Sensitive Marker

Abstract Chromogranins A, B, and C, proteins that are co-stored and co-released with peptides and amines, have been identified in a variety of neuroendocrine tissues, both normal and neoplastic. We examined the secretion of chromogranin A and chromogranin A + B by hormone-producing tumours in patients with endocrine pancreatic tumours, carcinoid tumours, pheochromocytomas, and small cell lung cancer. The radioimmunoassay determining the plasma concentrations of chromogranin A + B showed a greater sensitivity than that determining chromogranin A alone. All patients with endocrine pancreatic tumours, carcinoids, and pheochromocytomas had increased levels of chromograin A + B, whereas a small number of the patients (5/18 with endocrine pancreatic tumours and ⅓ with pheochromocytomas) had normal levels of chromogranin A. Also in immunocytochemical stainings, our polyclonal antiserum detecting both chromogranin A and B showed a greater sensitivity than other available antisera against chromogranin A, B and C. We have demonstrated that a polyclonal antiserum against a mixture of chromogranin A and B might be a more sensitive marker than chromogranin A alone for diagnosing neuroendocrine tumours. This is not surprising, since both chromogranins are widely distributed in neuroendocrine cells.

Immunoradiometric assay for alpha gamma- and gamma gamma-enolase (neuron-specific enolase), with use of monoclonal antibodies and magnetizable polymer particles.

1989 ◽  
Vol 35 (10) ◽  
pp. 2034-2038 ◽  
Author(s):  
E Paus ◽  
K Nustad
Keyword(s):  
Monoclonal Antibodies ◽  
Lung Carcinoma ◽  
Solid Phase ◽  
Small Cell Lung Carcinoma ◽  
Neuron Specific Enolase ◽  
Small Cell ◽  
Immunoradiometric Assay ◽  
Small Cell Lung ◽  
Cell Lung Carcinoma ◽  
Gamma Enolase

Abstract Monoclonal antibodies were raised against neuron-specific enolase, gamma gamma-enolase, and used in an immunoradiometric assay (IRMA), with mono-disperse magnetizable particles as the solid phase. The assay's sensitivity was 0.4 microgram/L and the interassay coefficient of variation was less than 5% in the working range from 0.4 to 170 micrograms/L. Compared with our radioimmunoassay based on polyclonal antibodies, the incubation time is shorter, and precision and sensitivity are improved. The IRMA also improved detection of neuron-specific enolase in sera from patients with lung cancer without a concomitant change in measured enolase in the reference population. The better sensitivity of the IRMA results from its ability to measure alpha gamma- and gamma gamma-enolase with equal response. Ninety percent of the small-cell lung carcinoma patients (36 of 40) had increased values before treatment, compared with 7% of non-small-cell lung carcinoma patients (8 of 114).

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