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https://www.readbyqxmd.com/read/28805498/gingiskhan%C3%A2-protease-cleavage-allows-a-high-throughput-antibody-to-fab-conversion-enabling-direct-functional-assessment-during-lead-identification-of-human-monoclonal-and-bispecific-igg1-antibodies
#1
Jörg Moelleken, Manuel Endesfelder, Christian Gassner, Sabine Lingke, Simone Tomaschek, Oksana Tyshchuk, Stefan Lorenz, Ulrike Reiff, Michael Mølhøj
The determination of the binding strength of immunoglobulins (IgGs) to targets can be influenced by avidity when the targets are soluble di- or multimeric proteins, or associated to cell surfaces, including surfaces introduced from heterogeneous assays. However, for the understanding of the contribution of a second drug-to-target binding site in molecular design, or for ranking of monovalent binders during lead identification, affinity-based assessment of the binding strength is required. Typically, monovalent binders like antigen-binding fragments (Fabs) are generated by proteolytic cleavage with papain, which often results in a combination of under- and over-digestion, and requires specific optimization and chromatographic purification of the desired Fabs...
August 14, 2017: MAbs
https://www.readbyqxmd.com/read/28800544/protease-loaded-permeation-enhancer-liposomes-for-treatment-of-skin-fibrosis-arisen-from-second-degree-burn
#2
Kamini Sahu, Monika Kaurav, Ravi Shankar Pandey
Cysteine protease (papain) is a plant derived enzyme and due to its collagenolytic activity has potential in fibrosis reduction. However, a major hurdle in its use as fibrosis reducing agent is to overcome stratum corneum skin barrier via topical application, owing to its hydrophilic and high molecular weight and protein nature which is prone to degradation. The aim of the present study was to develop a penetration enhancer incorporated drug delivery system, i.e. propylene glycol (PG) liposomes, loaded with papain for application in fibrosis therapy...
August 8, 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28799505/single-molecular-level-probing-of-structure-and-dynamics-of-papain-under-denaturation
#3
Bhaswati Sengupta, Apala Chaudhury, Nilimesh Das, Pratik Sen
INTRODUCTION: Papain is a cysteine protease enzyme present in papaya and known to help in digesting peptide. Thus the structure and function of the active site of papain is of interest. OBJECTIVE: The objective of present study is to unveil the overall structural transformation and the local structural change around the active site of papain as a function of chemical denaturant. METHODS: Papain has been tagged at Cys-25 with a thiol specific fluorescence probe N-(7-dimethylamino-4-methylcoumarin-3-yl) iodoacetamide (DACIA)...
August 11, 2017: Protein and Peptide Letters
https://www.readbyqxmd.com/read/28791512/purification-of-angiotensin-i-converting-enzyme-ace-inhibitory-peptides-from-casein-hydrolysate-by-imac-ni-2
#4
Shanguang Wu, Xuezhen Feng, Yuan Lu, Yuting Lu, Saisai Liu, Yuhong Tian
Casein proteins were hydrolyzed by papain to identify inhibitory peptides of angiotensin I-converting enzyme (ACE). The hydrolysate was fractionized by immobilized metal affinity chromatography (IMAC-Ni(2+)). The fraction with high ACE inhibitory activity was enriched and further chromatographed on a reverse-phase column to yield four fractions. Among the fractions, the L4 fraction exhibited the highest ACE inhibitory activity and was identified by sequence analysis as Trp-Tyr-Leu-His-Tyr-Ala (WYLHYA), with IC50 value of 16...
August 8, 2017: Amino Acids
https://www.readbyqxmd.com/read/28782582/development-and-in-vitro-characterization-of-a-papain-loaded-mucolytic-self-emulsifying-drug-delivery-system-sedds
#5
Christina Leichner, Claudia Menzel, Flavia Laffleur, Andreas Bernkop-Schnürch
The aim of the study was to create a self-emulsifying drug delivery system (SEDDS) with mucolytic properties based on incorporated papain for improved mucus permeation. In order to increase the lipophilicity of the enzyme and to dissolve it in SEDDS, hydrophobic ion pairing with sodium deoxycholate in a molar ratio of 20:1 (surfactant: enzyme) was performed. The yield of precipitated papain was 86.8±2.7% and the ion pair was loaded into the formulations to 1% (m/m). Suitable formulations were chosen according to their properties to dissolve the ion pair and characterized regarding droplet size and polydispersity index...
August 3, 2017: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/28771551/computational-investigation-of-conformational-variability-and-allostery-in-cathepsin-k-and-other-related-peptidases
#6
Marko Novinec
Allosteric targeting is progressively gaining ground as a strategy in drug design. Its success, however, depends on our knowledge of the investigated system. In the case of the papain-like cysteine peptidase cathepsin K, a major obstacle in our understanding of allostery is represented by the lack of observable conformational change at the active site. This makes it difficult to understand how binding of effectors at known allosteric sites translates into modified enzyme activity. Herein, we address this issue by a computational approach based on experimental data...
2017: PloS One
https://www.readbyqxmd.com/read/28754039/nanodiamond-chymotrypsin-and-nanodiamond-papain-conjugates-their-synthesis-and-activity-and-visualization-of-their-interaction-with-cells-using-optical-and-electron-microscopy
#7
Sergey A Golyshev, Anna K Berkovich, Ruslan Yu Yakovlev, Dmitry M Bystrov, Nikita M Ivanov, Galina N Balandina, Galina N Rudenskaya
Two novel conjugates of detonation nanodiamonds (dNDs) with the proteolytic enzymes chymotrypsin and papain were synthesized. The synthesis was performed via functionalization of the dNDs' surface with acidic/alkali treatment followed by carbodiimide-mediated protein binding. Covalent binding of the enzymes was confirmed by Fourier transform infrared spectrographic analysis and high-performance liquid chromatography (HPLC) amino acid analysis. HPLC also proved the preservation of the enzymes' composition during synthesis...
July 28, 2017: Biointerphases
https://www.readbyqxmd.com/read/28752264/papain-like-cysteine-protease-encoding-genes-in-rubber-hevea-brasiliensis-comparative-genomics-phylogenetic-and-transcriptional-profiling-analysis
#8
Zhi Zou, Guishui Xie, Lifu Yang
43 HbPLCPs representing nine subfamilies or 20 orthologous groups were found in rubber, where paralogs were resulted from the recent WGD and local duplication. Several senescence-associated genes were also identified. Papain-like cysteine proteases (PLCPs) comprise a large family of proteolytic enzymes involved in plant growth and development, seed germination, organ senescence, immunity, and stress response. Despite their importance and the extensive research in the model plant Arabidopsis thaliana, little information is available on rubber tree (Hevea brasiliensis), a rubber-producing plant of the Euphorbiaceae family...
July 27, 2017: Planta
https://www.readbyqxmd.com/read/28731389/protease-hydrolysates-of-filefish-thamnaconus-modestus-byproducts-effectively-inhibit-foodborne-pathogens
#9
Shaimaa Hatab, Mei-Ling Chen, Wenhua Miao, Jiheng Lin, Dandan Wu, Changyu Wang, Pengxiang Yuan, Shanggui Deng
In this study, novel antimicrobial peptides had been derived by enzymatic hydrolysis of filefish (Thamnaconus modestus) byproduct (HFBP). Different proteases, (papain [P], trypsin [T], neutrase [N], pepsin [PE], and the mixture I [PN] and mixture II [PT]) and different hydrolysis time (60, 120, 180, and 240 min), have been used to generate peptides with different lengths and amino acid sequences. The antimicrobial properties of HFBPs were tested, against Gram-positive and Gram-negative strains, using disc diffusion method...
July 21, 2017: Foodborne Pathogens and Disease
https://www.readbyqxmd.com/read/28730605/contributions-of-interleukin-33-and-tslp-in-a-papain-soaked-contact-lens-induced-mouse-conjunctival-inflammation-model
#10
Jobu Sugita, Yosuke Asada, Waka Ishida, Satoshi Iwamoto, Katsuko Sudo, Hajime Suto, Toru Matsunaga, Ken Fukuda, Atsuki Fukushima, Norihiko Yokoi, Tatsukuni Ohno, Miyuki Azuma, Nobuyuki Ebihara, Hirohisa Saito, Masato Kubo, Susumu Nakae, Akira Matsuda
INTRODUCTION: Pathological changes of severe chronic allergic conjunctivitis are driven not only via acquired immunity but also via innate immunity. Type 2 immune response-initiating cytokines may play some roles as innate immunity-dependent components of the ocular surface inflammation. To investigate the involvement of type 2 immune response-initiating cytokines in innate immunity-dependent, papain-induced conjunctival inflammation model using IL-25-, IL-33-, and TSLP receptor (TSLPR)-knockout (KO) mice with reference to basophils and ILC2...
July 20, 2017: Immunity, Inflammation and Disease
https://www.readbyqxmd.com/read/28720947/reduction-of-whey-protein-concentrate-antigenicity-by-using-a-combined-enzymatic-digestion-and-ultrafiltration-approach
#11
Laura Quintieri, Linda Monaci, Federico Baruzzi, Maria Gabriella Giuffrida, Silvia de Candia, Leonardo Caputo
The global interest in saving food resources is leading to recycle wasted-food materials to extract useful nutrients. In dairy industry, the recycling of whey proteins determines their utilization in the healthy-addressed foods, which, however, can cause immunological responses in allergic subjects. In this work, a whey protein concentrate (WPC) was alternatively hydrolyzed with pepsin, papain, trypsin and rennin in order to attenuate or abolish the β-lactoglobulin (BLG) antigenicity. The electrophoretic profiles of both pepsin and papain WPC hydrolysates proved the disappearance of the BLG band, even though a slight antigenicity was still found by ELISA...
June 2017: Journal of Food Science and Technology
https://www.readbyqxmd.com/read/28711271/identification-of-dipeptidyl-peptidase-iv-inhibitory-peptides-from-mare-whey-protein-hydrolysates
#12
J J Song, Q Wang, M Du, X M Ji, X Y Mao
Inhibition of dipeptidyl peptidase-IV (DPP-IV) activity is a promising strategy for treatment of type 2 diabetes. In the current study, DPP-IV inhibitory peptides were identified from mare whey protein hydrolysates obtained by papain. The results showed that all the mare whey protein hydrolysates obtained at various hydrolysis durations possessed more potent DPP-IV inhibitory activity compared with intact whey protein. The 4-h hydrolysates showed the greatest DPP-IV inhibitory activity with half-maximal inhibitory concentration of 0...
September 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/28706952/development-of-an-economical-method-to-reduce-the-extractable-latex-protein-levels-in-finished-dipped-rubber-products
#13
Ambegoda Liyanage Harini Amalka Perera, Bulathsinhalage Gayani Kanchana Perera
Natural rubber latex (NRL) allergy is caused by the extractable latex proteins in dipped rubber products. It is a major concern for the consumers who are sensitive to the allergenic extractable proteins (EP) in products such as NRL gloves. Objective of this research was to develop an economical method to reduce the EP in finished dipped NRL products. In order to reduce the EP levels, two natural proteases, bromelain from pineapple and papain from papaya, were extracted and partially purified using (NH4)2SO4...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28702797/purification-and-antibacterial-mechanism-of-fish-borne-bacteriocin-and-its-application-in-shrimp-penaeus-vannamei-for-inhibiting-vibrio-parahaemolyticus
#14
Xinran Lv, Jingfang Du, Yu Jie, Bolin Zhang, Fengling Bai, Hongfei Zhao, Jianrong Li
Vibrio parahaemolyticus: is recognized as the main cause of gastroenteritis associated with consumption of seafood. Bacteriocin-producing Lactobacillus plantarum FGC-12 isolated from golden carp intestine had strong antibacterial activity toward V. parahaemolyticus. The fish-borne bacteriocin was purified by a three-step procedure consisting of ethyl acetate extraction, gel filtration chromatography and high performance liquid chromatography. Its molecular weight was estimated at 4.1 kDa using SDS-PAGE. The fish-borne bacteriocin reached the maximum production at stationary phase after 20 h...
August 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28697819/functional-characterization-of-single-domain-cystatin-like-cysteine-proteinase-inhibitors-expressed-by-the-trematode-fasciola-hepatica
#15
Martín Cancela, Ileana Corvo, Edileuza DA Silva, Aline Teichmann, Leda Roche, Alvaro Díaz, José Fransisco Tort, Henrique B Ferreira, Arnaldo Zaha
Cystatins are small, phylogenetically conserved proteins that are tight-binding inhibitors of cysteine proteinases. The liver fluke Fasciola hepatica uses a diverse set of cysteine proteinases of the papain superfamily for host invasion, immune evasion and nutrition, but little is known about the regulation of these enzymes. The aim of this work is to characterize the cystatin repertoire of F. hepatica. For this purpose, we first surveyed the available sequence databases, identifying three different F. hepatica single-domain cystatins...
July 12, 2017: Parasitology
https://www.readbyqxmd.com/read/28690166/immune-response-of-anti-lectin-pjlec-antibody-in-freshwater-crab-paratelphusa-jacquemontii
#16
Maghil Denis, Sivakumar Mullaivanam Ramasamy, Karthigayani Thayappan, Arumugam Munusamy
Sialic acid specific lectin Pjlec isolated from serum of the freshwater crab Paratelphusa jacquemontii served as an antigen for the production of immunoglobulin (Ig) in Balb/c mice sera. Enzyme-linked immunosorbent assay (ELISA) of mice anti-sera with Pjlec lectin affirmed the induction and production of antibody. Anti-Pjlec antibody was isolated from the antisera of mice by Protein A Sepharose affinity chromatography and checked for purity by immunoblot with lectin. Mass spectrometry (MS/MS) of papain digethe peptide sequence of antigen binding fragment (Fab) and fragment crystallizable (Fc)...
July 8, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28687334/a-cysteine-protease-cathepsin-z-from-disk-abalone-haliotis-discus-discus-genomic-characterization-and-transcriptional-profiling-during-bacterial-infections
#17
G I Godahewa, N C N Perera, Sukkyoung Lee, Myoung-Jin Kim, Jehee Lee
Cathepsin Z (CTSZ) is lysosomal cysteine protease of the papain superfamily. It participates in the host immune defense via phagocytosis, signal transduction, cell-cell communication, proliferation, and migration of immune cells such as monocytes, macrophages, and dendritic cells. Hence, CTSZ is also acknowledged as an acute-phase protein in host immunity. In this study, we sought to identify the CTSZ homolog from disk abalone (AbCTSZ) and characterize it at the molecular, genomic, and transcriptional levels...
July 4, 2017: Gene
https://www.readbyqxmd.com/read/28683142/the-plasmodium-falciparum-pseudoprotease-sera5-regulates-the-kinetics-and-efficiency-of-malaria-parasite-egress-from-host-erythrocytes
#18
Christine R Collins, Fiona Hackett, Jonathan Atid, Michele Ser Ying Tan, Michael J Blackman
Egress of the malaria parasite Plasmodium falciparum from its host red blood cell is a rapid, highly regulated event that is essential for maintenance and completion of the parasite life cycle. Egress is protease-dependent and is temporally associated with extensive proteolytic modification of parasite proteins, including a family of papain-like proteins called SERA that are expressed in the parasite parasitophorous vacuole. Previous work has shown that the most abundant SERA, SERA5, plays an important but non-enzymatic role in asexual blood stages...
July 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28666223/protocols-for-the-analytical-characterization-of-therapeutic-monoclonal-antibodies-ii-enzymatic-and-chemical-sample-preparation
#19
Balazs Bobaly, Valentina D'Atri, Alexandre Goyon, Olivier Colas, Alain Beck, Szabolcs Fekete, Davy Guillarme
The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. These approaches make the sample more suitable for chromatographic and mass spectrometric analysis...
June 19, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/28666163/macroporous-monoliths-for-biodegradation-study-of-polymer-particles-considered-as-drug-delivery-systems
#20
M V Volokitina, V A Korzhikov-Vlakh, T B Tennikova, E G Korzhikova-Vlakh
Nanostructures based on biodegradable polymers are often considered as drug delivery systems. The properties of these nanomaterails towards in vitro biodegradation are very important and usually are studied using the model physiological conditions. In this work the novel approach based on application of monolithic immobilized enzyme reactors (IMERs) as the systems for biodegradation study of the nanoobjects of different nature and morphology was suggested. Rigid nanospheres based on poly(lactic acid) and self-assembled nanoobjects formed from block-copolymer of glutamic acid and phenylalanine were applied as model nanomaterials...
June 20, 2017: Journal of Pharmaceutical and Biomedical Analysis
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