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Preparation and Identification of Monoclonal Antibody against fluorescein isothiocyanate

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Preparation and Identification of Monoclonal Antibody agains...

A new quality control method for lateral flow assay

第十二届全国免疫学学术大会

作者： niu yan Li Jinge Zhang Gaiping zhengzhou university Henan agricultural university

fluorescein isothiocyanate(FITC) is a hapten with no immunogenic *** order to generate immunogenicity,the artificial antigens,FITC-BSA and FITC-OVA,were *** BALB/c mice were immunized three times with *** the polyethylene glycol method,splenocytes from the immunized mice were fused with mouse myeloma cells SP2/*** subcloning,a strong positive 2 A7 hybridoma clone was obtained,which was used to generate ascites *** analysis showed that most of the 2 A7 monoclonal antibody(mAb) belonged to the immunoglobulin Gl(Kappa-light chain) *** titer of 2 A7 mAb was 2.048×10and the FITC concentration that caused50%inhibition(IC50) of binding of 2 A7 mAb to FITC-OVA was 1.37 μg/*** results of cross-reactivity showed that2 A7 mAb was highly specific to FITC,and the affinity of 2 A7 mAb was 1.72×10 L/mol,suggesting that the 2 A7 mAb was high affinity to *** conclusion,the anti-FITC mAb produced in the current study provides a useful tool for the effective detection of FITC in various types of immunoassays.

关键词： fluorescein isothiocyanate enzyme-linked immunosorbent assay monoclonal antibody

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Chinese Chemical Letters 2018年第12期29卷 1853-1856页

作者： Li Huang Di Zhang Lirong Jiao Enben Su Nongyue He State Key Laboratory of Bioelectronics School of Biological Science and Medical Engineering Southeast University National Demonstration Center for Experimental Biomedical Engineering Education Southeast University Getein Biotechnology Co. Ltd.

Precision and repeatability are challenging issues in point of care testing(POCT) analysis. Herein, we proposed a lateral flow assay(LFA) based on internal quality control microspheres to realize the accurate diagnosis of HbAlc in human body. fluorescein cy5 decorated microspheres are used as labels for HbAlc detection, and BSA-fluorescein isothiocyanate(FITC) decorated microspheres are used as internal quality control labels. One test line was employed in the strip for the detection of glycosylated hemoglobin(HbAlc). This method can eliminate the interference of environmental factors(temperature, humidity,etc.) to LFA in the process of chromatography, and improve the precision and accuracy of HbAlc *** CV for detection of low concentration HbAlc was 1.05%, and the CV for detection of high concentration HbAlc was 0.69%. We envision the method to have great prospect in in vitro diagnosis(IVD).

关键词： Point of care testing Lateral flow assay HbA1c fluorescein isothiocyanate Precision

Preparation, Characterization and Pharmacokinetics of Fluorescence Labeled Propylene Glycol Alginate Sodium Sulfate

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Journal of Ocean University of China 2014年第4期13卷 683-690页

作者： LI Pengli LI Chunxia XUE Yiting ZHANG Yang LIU Hongbing ZHAO Xia YU Guangli GUAN Huashi Key Laboratory of Marine Drugs of Ministry of Education Provincial Key Laboratory of Glycoscience and Glycotechnology of Shandong School of Medicine and Pharmacy Ocean University of China

A rapid and sensitive fluorescence labeling method was developed and validated for the microanalysis of a sulfated polysaccharide drug,namely propylene glycol alginate sodium sulfate(PSS), in rat plasma. fluorescein isothiocyanate(FITC) was selected to label PSS, and 1, 6-diaminohexane was used to link PSS and FITC in order to prepare FITC-labeled PSS(F-PSS) through a reductive amination reaction. F-PSS was identified by UV-Vis, FT-IR and 1H-NMR spectrum. The cell stability and cytotoxicity of F-PSS were tested in Madin-Darby canine kidney(MDCK) cells. The results indicated that the labeling efficiency of F-PSS was 0.522% ± 0.0248% and the absolute bioavailability was 8.39%. F-PSS was stable in MDCK cells without obvious cytotoxicity. The method was sensitive and reliable; it showed a good linearity, precision, recovery and stability. The FITC labeling method can be applied to investigating the absorption and metabolism of PSS and other polysaccharides in biological samples.

关键词： propylene glycol alginate sodium sulfate fluorescence labeling fluorescein isothiocyanate pharmacokinetics

Pharmacokinetics and Biodegradation Performance of a Hydroxypropyl Chitosan Derivative

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Journal of Ocean University of China 2015年第5期14卷 888-896页

作者： SHAO Kai HAN Baoqin DONG Wen SONG Fulai LIU Weizhi LIU Wanshun College of Marine Life Science Ocean University of China

Hydroxypropyl chitosan(HP-chitosan) has been shown to have promising applications in a wide range of areas due to its biocompatibility, biodegradability and various biological activities, especially in the biomedical and pharmaceutical fields. However, it is not yet known about its pharmacokinetics and biodegradation performance, which are crucial for its clinical applications. In order to lay a foundation for its further applications and exploitations, here we carried out fluorescence intensity and GPC analyses to determine the pharmacokinetics mode of fluorescein isothiocyanate-labeled HP-chitosan(FITC-HP-chitosan) and its biodegradability. The results showed that after intraperitoneal administration at a dose of 10 mg per rat, FITC-HP-chitosan could be absorbed rapidly and distributed to liver, kidney and spleen through blood. It was indicated that FITC-HP-chitosan could be utilized effectively, and 88.47% of the FITC-HP-chitosan could be excreted by urine within 11 days with a molecular weight less than 10 k Da. Moreover, our data indicated that there was an obvious degradation process occurred in liver(< 10 k Da at 24 h). In summary, HP-chitosan has excellent bioavailability and biodegradability, suggesting the potential applications of hydroxypropyl-modified chitosan as materials in drug delivery, tissue engineering and biomedical area.

关键词： hydroxypropyl chitosan fluorescein isothiocyanate pharmacokinetics biodegradation performance biomedicalmaterial

Pharmacokinetics and Biodegradation of Chitosan in Rats

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Journal of Ocean University of China 2015年第5期14卷 897-904页

作者： LI Hui JIANG Zhiwen HAN Baoqin NIU Shuyi DONG Wen LIU Wanshun College of Marine Life Sciences Ocean University of China

Chitosan, an excellent biomedical material, has received a widespread in vivo application. In contrast, its metabolism and distribution once being implanted were less documented. In this study, the pharmacokinetics and biodegradation of fluorescein isothiocyanate(FITC) labeled and muscle implantation administrated chitosan in rats were investigated with fluorescence spectrophotometry, histological assay and gel chromatography. After implantation, chitosan was degraded gradually during its distribution to diverse organs. Among the tested organs, liver and kidney were found to be the first two highest in chitosan content, which was followed by heart, brain and spleen. Urinary excretion was believed to be the major pathway of chitosan elimination, yet 80% of chitosan administered to rats was not trackable in their urine. This indicated that the majority of chitosan was degraded in tissues. In average, the molecular weight of the degradation products of chitosan in diverse organs and urine was found to be <65 k Da. This further confirmed the in vivo degradation of chitosan. Our findings provided new evidences for the intensive and safe application of chitosan as a biomedical material.

关键词： chitosan fluorescein isothiocyanate pharmacokinetics biodegradation fluorescence spectrophotometry muscle implantation