Anti-His tag antibody [Biotin]

Sugarcane and energy cane (Saccharumspp. hybrids) are ideal for plant-based production of recombinant proteins because their high resource-use efficiency, rapid growth and efficient photosynthesis enable extensive biomass production and protein accumulation at a cost-effective scale. Here, we aimed to develop these species as efficient platforms to produce recombinantGalanthus nivalisL. (snowdrop) agglutinin (GNA), a monocot-bulb mannose-specific lectin with potent antiviral, antifungal and antitumor activities. Initially, GNA levels of 0.04% and 0.3% total soluble protein (TSP) (0.3 and 3.8 mg kg(-1)tissue) were recovered from the culms and leaves, respectively, of sugarcane lines expressing recombinantGNAunder the control of the constitutive maizeubiquitin 1(Ubi) promoter. Co-expression of recombinant GNA from stacked multiple promoters (pUbiand culm-regulated promoters from sugarcanedirigent5-1andSugarcane bacilliform virus) on separate expression vectors increased GNA yields up to 42.3-fold (1.8% TSP or 12.7 mg kg(-1)tissue) and 7.7-fold (2.3% TSP or 29.3 mg kg(-1)tissue) in sugarcane and energy cane lines, respectively. Moreover, inducing promoter activity in the leaves of GNA transgenic lines with stress-regulated hormones increased GNA accumulation to 2.7% TSP (37.2 mg kg(-1)tissue). Purification by mannose-agarose affinity chromatography yielded a functional sugarcane recombinant GNA with binding substrate specificity similar to that of native snowdrop-bulb GNA, as shown by enzyme-linked lectin and mannose-binding inhibition assays. The size and molecular weight of recombinant GNA were identical to those of native GNA, as determined by size-exclusion chromatography and MALDI-TOF mass spectrometry. This work demonstrates the feasibility of producing recombinant GNA at high levels inSaccharumspecies, with the long-term goal of using it as a broad-spectrum antiviral carrier molecule for hemopurifiers and in related therapeutic applications.

Acinetobacter baumannii has gained attention for years as a significant clinical problem due to the increase of antibiotic-resistant strain. Indeed,A. baumannii OmpA is one of the highly conserved membrane proteins among Gram-negative bacteria that has multiple roles in interacting with the host during infection, thereby representing an effective target for the development of novel antibacterial or vaccination therapies. Nowadays, finding suitable epitope-specific antigens inside the conserved proteins such as OmpA is a promising method for successful vaccination programs. Therefore, in the present study, the coding sequence of the 240 to 356 amino acid residues of A. baumannii OmpA (AbOmpA240-356) was cloned into the vector pET-28a and purified using nickel affinity chromatography. In addition, the anti-His tag antibody is used to validate its production. This system of protein expression and purification maybe useful for further characterization of AbOmpA240-356 protein fraction. Therefore, this study can lead to the introduction of suitable candidates for the development of an effective vaccine based on OmpA against this bacterium for further analysis.