Development of a nanobody-based immunoassay for the sensitive detection of fibrinogen-like protein 1.

Immune checkpoint inhibition is an important strategy in cancer therapy. Blockade of CTLA-4 and PD-1/PD-L1 is well developed in clinical practice. In the last few years, LAG-3 has received much interest as an emerging novel target in immunotherapy. It was recently reported that FGL1 is a major ligand of LAG-3, which is normally secreted by the liver but is upregulated in several human cancers. FGL1 is a crucial biomarker and target for cancer immunotherapy. As the efficacy of immunotherapy is limited to specific types of patients, the subset of patients needs to be selected appropriately to receive precise treatment according to different biomarkers. To date, there is no test to accurately assess FGL1 expression levels. Nanobodies have some outstanding features, such as high stability, solubility and affinity for diagnostic and therapeutic applications. Here, we report the development and validation of a rapid, sensitive, and cost-effective nanobody-based immunoassay for the detection of FGL1 in human serum. In this study, human FGL1 recombinant protein was expressed and purified for the first time as an immunized antigen. Then, we constructed a nanobody phage display library and screened several nanobodies that bind FGL1 with high affinity. We selected two nanobodies targeting different epitopes of FGL1, one as a capture and the other conjugated with HRP as a probe. The double nanobody-based sandwich ELISA to detect the concentration of FGL1 showed a good response relationship in the range of 15.625-2000 ng/mL, and the recoveries from the spiked sample were in the range of 78% and 100%. This assay could be used as a potential approach for evaluating FGL1 expression for patient stratification and for predicting the therapeutic efficacy of targeting the LAG3/FGL1 axis.

A conductive sodium alginate and carboxymethyl chitosan hydrogel doped with polypyrrole for peripheral nerve regeneration.

Polymer materials with electrically conductive properties have good applications in their respective fields because of their special properties. However, they usually exhibited poor mechanical properties and biocompatibility. In this work, we present a simple approach to prepare conductive sodium alginate (SA) and carboxymethyl chitosan (CMCS) polymer hydrogels (SA/CMCS/PPy) that can provide sufficient help for peripheral nerve regeneration. SA/CMCS hydrogel was cross-linked by calcium ions provided by the sustained release system consisting of d-glucono-δ-lactone (GDL) and superfine calcium carbonate (CaCO3), and the conductivity of the hydrogel was provided by doped with polypyrrole (PPy). Gelation time, swelling ratio, porosity and Young's modulus of the conductive SA/CMCS/PPy hydrogel were adjusted by polypyrrole content, and the conductivity of it was within 2.41 × 10-5 to 8.03 × 10-3 S cm-1. The advantages of conductive hydrogels in cell growth were verified by controlling electrical stimulation of cell experiments, and the hydrogels were also used as a filling material for the nerve conduit in animal experiments. The SA/CMCS/PPy conductive hydrogel showed good biocompatibility and repair features as a bioactive biomaterial, we expect this conductive hydrogel will have a good potential in the neural tissue engineering.

[Identification on Xanthium sibiricum from different habitats by FTIR fingerprint patterns].

OBJECTIVE: To establish an FTIR method to identify Xanthium sibiricum from different habitats. METHODS: FTIR spectra of Xanthium sibiricum from different habitats were analyzed,and the similarity of different fingerprint spectra and the chemical pattern recognition were calculated and analyzed according to the wave numbers of peaks. RESULTS: Different FTIR spectra of 10 different habitats of Xanthium sibiricum were obtained,and the similarities were all above 0. 96. CONCLUSION: This method can be used for identification on Xanthium sibiricum from different habitats. The results of similarity calculation and chemical pattern recognition further prove the feasibility of this method.

[Identification on Solanum lyratum by FT-IR fingerprint patterns and similarity degree of different fingerprint patterns].

OBJECTIVE: To provide a FT-IR new method to identify different habitats of Solanum lyratum. METHODS: Analyzed FT-IR patterns of Solanum lyratum from different habitats, and the similarity of different fingerprint patterns was calculated and analyzed according to the wave numbers of peaks searched. RESULTS: Obtained the different FT-IR pattern of 5 different habitats of Solanum lyraturn. CONCLUSION: This method can be used for identifications on different habitats of Solanum lyratum. The results of similarity calculation further prove the feasibility of this method.

Preparation, characterization and in vitro release study of a glutathione-dependent polymeric prodrug Cis-3-(9H-purin-6-ylthio)-acrylic acid-graft-carboxymethyl chitosan.

In this work, an amphiphilic polymeric prodrug Cis-3-(9H-purin-6-ylthio)-acrylic acid-graft-carboxymethyl chitosan (PTA-g-CMCS) was designed and synthesized. In aqueous solution, this grafted polymer can self-assemble into spherical micelles with a size ranging from 104 to 285 nm and zeta potential ranging from -12.3 to -20.1 mV. For the release study, less than 24% of 6-Mercaptopurine (6-MP) was released from PTA-g-CMCS1 in the media containing 2 and 100 µM glutathione (GSH), whereas 37%, 54% and 75% of 6-MP was released from the media with GSH of 1, 2 and 10mM, respectively. Besides, pH and drug content of the polymeric prodrug only presented slight influence on the 6-MP release. MTT assay demonstrated that this system had higher inhibition ratio on HL-60 cells (human promyelocytic leukemia cells) in the presence of GSH and lower cytotoxicity on mouse fibroblast cell line (L929). Therefore, this nano-sized system is glutathione-dependent, and it can be employed as a potential carrier for the controlled release of 6-MP.

[Identification on Solanum lyratum by X-ray diffraction Fourier fingerprint and similarity analysis].

OBJECTIVE: To provide an X-ray diffraction (XRD) method for identifying different medicinal parts of Solanum lyratum. METHODS: Analyzed X-ray diffraction Fourier patterns of different parts of Solanum lyratum, and the similarity degree of different fingerprint was calculated and analyzed according to the position (2 theta value)of peaks searched. RESULTS: Different X-ray diffraction Fourier patterns of different medicinal parts of Solanum lyratum were obtained. CONCLUSION: This method can be used for identifying different medicinal parts of Solanum lyratum. The results of similarity calculation further proves the feasibility of this method.

[Optimization for formulation matrix proportion of xiangyu cataplasm by uniform design].

OBJECTIVE: To optimize the matrix formulation of cataplasm used to cure infantile diarrhea. METHODS: The optimum proportion of matrix for the preparation technology process of cataplasm was selected by uniform design and SPSS regression analysis. A check-up for adhibition , peeling strength, nonflowing, content of cream was founded. RESULTS: The best matrix's prescription gelatin: CMC-Na: PANA: kaolin: aluminum trichloride: citric acid: PVP K-30: PEG400: trimethylene glycol: tween-80 was 0.25 : 0.1 : 0.2 : 1.5 : 0.4 : 0.6 : 0.8 : 2 : 1 : 0.5. CONCLUSION: The preparation technique of cataplasm is feasible, and its quality is steerable, it is a safe and effective transdermal-drug delivery system.