Rapid production of therapeutic proteins using plant system

Plant molecular farming is simply defined as the production of proteins therapeutics (PT) in plants, which involves transient gene expression in plants and purification of expressed protein to a great scale for diagnosis, treatment and other applications.  This is therapid,economical, safe and reproducible approach for the production of PTas compared to bacterial and mammalian systems. Protein yield and post-translational modifications are the major roadblocks that can be overcome byhigh expression strategies includes over expression constructs, suitable plant host systems and glycoengineering of proteins. The inherent ability of ideally producing safe, functional protein is the most striking phenomenon recognized by the pharmaceutical industries and developed many therapeutic products within few weeks to meet escalating demands during pandemic/epidemic outbreaks recentl

Immunotherapy targeting isoDGR-protein damage extends lifespan in a mouse model of protein deamidation

\ua9 2023 The Authors. Published under the terms of the CC BY 4.0 license. Aging results from the accumulation of molecular damage that impairs normal biochemical processes. We previously reported that age-linked damage to amino acid sequence NGR (Asn-Gly-Arg) results in “gain-of-function” conformational switching to isoDGR (isoAsp-Gly-Arg). This integrin-binding motif activates leukocytes and promotes chronic inflammation, which are characteristic features of age-linked cardiovascular disorders. We now report that anti-isoDGR immunotherapy mitigates lifespan reduction of Pcmt1−/− mouse. We observed extensive accumulation of isoDGR and inflammatory cytokine expression in multiple tissues from Pcmt1−/− and naturally aged WT animals, which could also be induced via injection of isoDGR-modified plasma proteins or synthetic peptides into young WT animals. However, weekly injection of anti-isoDGR mAb (1 mg/kg) was sufficient to significantly reduce isoDGR-protein levels in body tissues, decreased pro-inflammatory cytokine concentrations in blood plasma, improved cognition/coordination metrics, and extended the average lifespan of Pcmt1−/− mice. Mechanistically, isoDGR-mAb mediated immune clearance of damaged isoDGR-proteins via antibody-dependent cellular phagocytosis (ADCP). These results indicate that immunotherapy targeting age-linked protein damage may represent an effective intervention strategy in a range of human degenerative disorders

Isolation and partial characterization of the active metabolite of ascidian, Polyclinum madrasensis from the Palk Bay Region, Southeast coast of India

Ascidians are rich source of bioactive agents which could be used for novel antimicrobial drugs. The present investigation inspects the antibacterial potential of ascidian, Polyclinum madrasensis collected from Mandapam, the Palk Bay region, Southeast coast of India. The crude extracts were tested for inhibition of bacterial growth against human pathogens.  Antibacterial assay was carried out by agar well diffusion method. The maximum inhibition zone (12.0±0.5 mm) was observed against the Staphylococcus aureus in crude methanol extract. The consequent zone of 6.5±0.1 mm was observed against S. aureus in ethanol extract and minimum inhibition zone (3.2±0.5 mm) was noticed with Pseudomonas aeruginosa. Molecular weight of tissue protein was determined through sodium dodecyl sulphate polyacryalamide gel electrophoresis (SDS-PAGE) and active metabolites were characterized by Fourier Transform Infrared spectroscopy (FTIR) analysis. The protein bands were at 36.5, 20.5, and 10.5 kDa, in SDS-PAGE and O-H stretch carboxylic acid compounds identified at the peak 3533.59 cm-1. It could be concluded from the present study that crude extract of the ascidian, P. madrasensis has potential antimicrobial effect against human pathogens.Keywards: Polyclinum madrasensis, bioactive compounds, Fourier Transform Infrared spectroscopy (FT-IR), sodium dodecyl sulphate polyacryalamide gel electrophoresis (SDS-PAGE).African Journal of Biotechnology, Vol 13(48) 4471-447

Response surface method to optimize the low cost medium for protease production using anchovy meal from ascidian associated Bacillus sp. GA CAS10

A protease producing Bacillus sp. GA CAS10 was isolated from ascidian Phallusia arabica, Tuticorin, Southeast coast of India. Response surface methodology was employed for the optimization of different nutritional and physical factors for the production of protease. Plackett-Burman method was applied to identify important factors (anchovy waste, KH2PO4, NaCl and temperature) influencing protease production. Further optimization was done by response surface methodology using central composite design. Under the optimized conditions by central composite design, the protease experimental yield (842.102 U/ml) closely matched the predicted yield by the statistical model (830.307 U/ml) with R2 = 99.94%. The time course of protease production was increased using the RSM optimized medium (856.29 U/ml) (anchovy waste 20.50 g/l, KH2PO4 3.06 g/l, NaCl 42.91 g/l, temperature 43.36°C, 42 h and pH 9) compared with the un-optimized basal medium (267.33 U/ml). The improvement of protease production by microbial conversion of anchovy waste suggested its potential utilization to generate high value added products using cheap carbon and nitrogen substrates.  Keywords: Protease, anchovy waste, statistical optimization, ascidian associated bacteria.African Journal of Biotechnology Vol 13(27) 2741-274

An investigation on the growth and propitiates of KDP admixtured ADP single crystals

ADP:KDP (85:15) mixed crystal was grown using slow cooling technique. The crystalline perfection of the grown crystal was studied using HRXRD curve. The homogeneity of KDP along the length of mixed crystal was analysed. Low Urbach energy value of the grown crystal indicates good crystallinity of the mixed crystals. Etch pit density was found using chemical etching studies. Vickers hardness studies indicate that the quality of the mixed crystal is more stable compared to ADP. The photoconductive nature and piezoelectric coefficient of the crystal were found. The laser stability of the crystal was observed at 532?nm using Nd:YAG laser

Immunotherapy targeting isoDGR-protein damage extends lifespan in a mouse model of protein deamidation

Aging results from the accumulation of molecular damage that impairs normal biochemical processes. We previously reported that age-linked damage to amino acid sequence NGR (Asn-Gly-Arg) results in "gain-of-function" conformational switching to isoDGR (isoAsp-Gly-Arg). This integrin-binding motif activates leukocytes and promotes chronic inflammation, which are characteristic features of age-linked cardiovascular disorders. We now report that anti-isoDGR immunotherapy mitigates lifespan reduction of Pcmt1-/- mouse. We observed extensive accumulation of isoDGR and inflammatory cytokine expression in multiple tissues from Pcmt1-/- and naturally aged WT animals, which could also be induced via injection of isoDGR-modified plasma proteins or synthetic peptides into young WT animals. However, weekly injection of anti-isoDGR mAb (1 mg/kg) was sufficient to significantly reduce isoDGR-protein levels in body tissues, decreased pro-inflammatory cytokine concentrations in blood plasma, improved cognition/coordination metrics, and extended the average lifespan of Pcmt1-/- mice. Mechanistically, isoDGR-mAb mediated immune clearance of damaged isoDGR-proteins via antibody-dependent cellular phagocytosis (ADCP). These results indicate that immunotherapy targeting age-linked protein damage may represent an effective intervention strategy in a range of human degenerative disorders