ABSTRACT
First identified as a high-affinity kinase-deficient receptor for class-3 semaphorins and vascular endothelial growth factor (VEGF) families, Neuropilin2 (NRP2) is a transmembrane non-tyrosine-kinase glycoprotein that has a vital function in neuronal patterning. Furthermore, NRP2 expression is often upregulated in cancer tissues and correlated with poor prognosis. In the present study, we report the establishment of a monoclonal antibody specific for NRP2b1b2 domain (NRP2 MAb) through hybridoma method. NRP2 MAb is measured to have a titer of 5.12 × 10(5) against NRP2b1b2 in indirect ELISA. Western blotting, flow cytometry, and immunofluorescence analysis indicate that NRP2 MAb can combine full-length NRP2 in LoVo and SW480 cells. Besides helping further understand NRP2-related pathological mechanisms and cell-signaling pathways, NRP2 MAb may act as a therapeutic agent for cancer in the future.
Subject(s)
Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/immunology , Neuropilin-2/immunology , Animals , Cells, Cultured , Enzyme-Linked Immunosorbent Assay/methods , Female , Fluorescent Antibody Technique/methods , Hybridomas/immunology , Membrane Proteins/chemistry , Membrane Proteins/immunology , Mice , Mice, Inbred BALB C , Protein Structure, Tertiary , Signal Transduction/immunology , Up-Regulation/immunology , Vascular Endothelial Growth Factor A/immunologyABSTRACT
Dextran as anti-nutritional factor is usually a result of bacteria activity and has associated serial problems during the process stream in the sugar industry and in medical therapy. A sensitive method is expected to detect dextran quantitatively. Here we generated four monoclonal antibodies (MAbs) against dextran using dextran T40 conjugated with bovine serum albumin (BSA) as immunogen in our lab following hybridoma protocol. Through pairwise, an MAb named D24 was determined to be conjugated with horseradish peroxidase (HRP) and was used in the establishment of a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) method for determination of dextran, in which MAb D9 was chosen as a capture antibody. The detection limit and working scope of the developed sandwich ELISA method were 3.9 ng/mL and 7.8-500 ng/mL with a correlation coefficient of 0.9909. In addition, the cross-reaction assay demonstrated that the method possessed high specificity with no significant cross-reaction with dextran-related substances, and the recovery rate ranged from 96.35 to 102.00%, with coefficient of variation ranging from 1.58 to 6.94%. These results indicated that we developed a detection system of MAb-based sandwich ELISA to measure dextran and this system should be a potential tool to determine dextran levels.
Subject(s)
Antibodies, Monoclonal/immunology , Dextrans/immunology , Enzyme-Linked Immunosorbent Assay/methods , Animals , Cell Line , Cross Reactions/immunology , Horseradish Peroxidase/immunology , Hybridomas/immunology , Limit of Detection , Mice , Sensitivity and SpecificityABSTRACT
In the title compound, [Ag(C(7)H(5)O(2))(C(5)H(6)N(2))(2)], the Ag(I) atom is tricoordinated by two independent pyridine N atoms and one benzoate O atom in a nearly planar geometry. An intramolecular N-H.O hydrogen bond forms an S(8) graph ring. The packing is built from molecular layers stabilized by two types of N-H.O hydrogen bond. Intermolecular Ag.N and intramolecular Ag.O contacts were also observed, together with three weak intermolecular C-H.pi interactions.
ABSTRACT
In the ternary title compound, catena-poly[[silver(I)-mu-ethylenediamine-kappa(2)N:N'] 3-nitrobenzoate monohydrate], [[Ag(C(2)H(8)N(2))](C(7)H(4)NO(4)) x H(2)O](n), the Ag atom is bicoordinated in a linear configuration by two different N atoms from two symmetry-related ethylenediamine ligands, thus giving linear polymeric chains with an [-Ag-N-C-C-N-](n) backbone running parallel to the a axis. In the crystal packing, these linear chains are interconnected by N-H...O and O-H...O hydrogen bonds to form layers parallel to the ab plane.
