A5035113807- Monoclonal and Polyclonal Antibodies Research
- Viral Infectious Diseases and Gene Expression in Insects
- Protein purification and stability
- Mast cells and histamine
- Asthma and respiratory diseases
- Allergic Rhinitis and Sensitization
- Food Allergy and Anaphylaxis Research
- Click Chemistry and Applications
- Glycosylation and Glycoproteins Research
- Nanoparticle-Based Drug Delivery
- bioluminescence and chemiluminescence research
- thermodynamics and calorimetric analyses
- Medical Research and Treatments
- RNA Interference and Gene Delivery
- Blood groups and transfusion
- Graphene and Nanomaterials Applications
- Synthesis and properties of polymers
- CRISPR and Genetic Engineering
- Chemical Synthesis and Analysis
- CAR-T cell therapy research
- Nanofabrication and Lithography Techniques
- Chemotherapy-induced organ toxicity mitigation
- Pancreatic function and diabetes
- Metal complexes synthesis and properties
- Virus-based gene therapy research
AstraZeneca (United States)
2020-2024
AstraZeneca (Netherlands)
2024
MacroGenics (United States)
2017
University of Notre Dame
2011-2014
Harvard University
2010
Cisplatin is a first line chemotherapy for most types of cancer. However, its use dose-limited due to severe nephrotoxicity. Here we report the rational engineering novel nanoplatinate inspired by mechanisms underlying cisplatin bioactivation. We engineered polymer, glucosamine-functionalized polyisobutylene-maleic acid, where platinum (Pt) can be complexed monomeric units using monocarboxylato and an O → Pt coordinate bond. show that at unique polymer ratio, this complex self-assembles into...
Abstract A key goal in process development for antibodies is to increase productivity while maintaining or improving product quality. During of an antibody, titers were increased from 4 10 g/L simultaneously decreasing aggregates. Process involved optimization media and feed formulations, strategy, parameters including pH temperature. To better understand how CHO cells respond changes, the changes implemented a stepwise manner. The first change was formulation, second medium, third...
Antibody disulfide bond reduction during monoclonal antibody (mAb) production is a phenomenon that has been attributed to the reducing enzymes from CHO cells acting on mAb harvest process. However, impact of downstream purification process not studied. During an IgG2 mAb, was observed in harvested cell culture fluid (HCCF), resulting high fragment levels. In addition, aggregate levels increased low pH treatment step A correlation between level free thiol HCCF (as result reduction) and...
The conserved nucleotide binding site (NBS), found within the Fab variable domain of antibodies, remains a not-so-widely known and underutilized site. Here we describe novel affinity chromatography method that utilizes NBS as target for selectively purifying antibodies from complex mixtures. column was prepared by coupling indole butyric acid (IBA), which has monovalent with K(d) ranging between 1 8 μM, to ToyoPearl resin resulting in targeting (NBS(IBA)). proof-of-concept studies performed...
Abstract One major challenge observed for the expression of therapeutic bispecific antibodies (BisAbs) is high product aggregates. Aggregates increase risk immune responses in patients and therefore must be removed at expense purification yields. BisAbs contain engineered disulfide bonds, which have been demonstrated to form aggregates, if mispaired. However, underlying intracellular mechanisms leading aggregate formation remain unknown. We demonstrate that impaired glutathione regulation...
Antibody interchain disulfide bond reduction during biopharmaceutical manufacturing has received increased attention since it was first reported in 2010. leads to loss of product and reduced stability. It is therefore critical understand the underlying mechanisms reduction. To date, thioredoxin system been as sole contributor antibody bioprocessing. In this work, we show that glutathione system, addition involved reducing molecules contributions two systems can vary depending upon cell...
Abstract The phenomenon of monoclonal antibody (mAb) interchain disulfide bond reduction during manufacturing processes continues to be a focus the biotechnology industry due potential for loss product, increased complexity purification processes, and reduced stability drug product. We hypothesized that can mitigated by controlling cell culture redox subsequently established threshold above which mAb remained intact below there were significant highly variable amounts mAb. Using this...
Liposome Extruder Purification (LEP) allows for the rapid purification of diverse liposome formulations using same extrusion apparatus employed during formation. The LEP process provides a means purifying functionalized liposomes from non-conjugated drug or protein contaminants with >93% recovery and contaminant removal in single step.
IgG4s are dynamic molecules that undergo a process called Fab-arm exchange. Disulfide bonds between heavy chains transiently reduced, resulting in half antibodies reform intact with other IgG4 antibodies. In vivo, therapeutic can recombine endogenous IgG4s, heterogeneous mixture of bispecific A related issue occur for any protein during manufacturing is interchain disulfide bond reduction. For this primarily results high levels half-mAb persist through purification processes. The S228P...
This study describes the design of a well-defined homotetravalent synthetic allergen (HTA) system to investigate effect hapten–IgE interactions on mast cell degranulation. A library DNP variants with varying affinities for IgEDNP was generated (Kd from 8.1 nM 9.2 μM), and 8 HTAs spanning this range were synthesized via conjugation each variant tetravalent scaffold. hapten Kd < 235 stimulated degranulation following bell-shaped dose response curve maximum occurring near Kd. ≥ failed stimulate...
The present paper describes the design of a HtTA (heterotetravalent allergen) as multi-component experimental system that enables an integrative approach to study mast cell degranulation. allows presentation two distinct haptens, each with valency 2, thereby better reflecting complexity natural allergens by displaying epitope heterogeneity and IgE antibody variability. Using design, synthetic HtTA-1 HtTA-2 were synthesized model combination epitope/IgE affinities. presented DNP...
Antibody disulfide bond (DSB) reduction during manufacturing processes is a widely observed phenomenon attributed to host cell reductases present in harvest culture fluid. Enzyme-induced antibody leads product fragments and aggregates that increase the impurity burden on purification process. The impact of bivalent bispecific antibodies (BisAbs), which are increasingly entering clinic, has yet be investigated. We focused reoxidation properties homologous library BisAb formats possess...
This Article describes an affinity-based precipitation method for the rapid and nonchromatographic purification of bivalently active monoclonal antibodies by combining selectivity affinity chromatography with simplicity salt-induced precipitation. procedure involves (i) proteins heavier than immunoglobulins ammonium sulfate; (ii) formation selective cyclic antibody complexes created binding to trivalent haptens specific antibody; (iii) membrane filtration solubilized pellet remove hapten...
This study describes a strategy where antibody selectivity for high antigen-density surfaces is enhanced by forming thermodynamically stable bicyclic complex. The complex was formed via multivalent interactions of the with synthetic trivalent mimotope at 3:2 molar ratio. Complex formation analyzed using dynamic light scattering and analytical ultracentrifugation, showing hydrodynamic radius ∼22 nm calculated molecular weight 397 kDa, depicting trimeric formation. has thermodynamic stability...
An affinity-based precipitation method was developed for the rapid and facile purification of bivalently active antibodies from complex biological solutions.
Current treatments for allergies include epinephrine and antihistamines, which treat the symptoms after an allergic response has taken place; steroids, result in local systemic immune suppression; IgE-depleting therapies, can be used only a narrow range of clinical IgE titers. The limitations current motivated design heterobivalent inhibitor (HBI) IgE-mediated responses that selectively inhibits allergen-IgE interactions, thereby preventing clustering mast cell degranulation. HBI was...
Abstract The global COVID‐19 pandemic ignited an unprecedented race to develop vaccines and antibody therapeutics. AstraZeneca's pursuit provide AZD7442 (EVUSHELD), two long‐acting, SARS‐CoV‐2 spike receptor binding domain‐specific neutralizing monoclonal antibodies, individuals at risk on highly accelerated timelines challenged our traditional ways of process development spurred the rapid adoption novel approaches. Conventional upstream processes were replaced by agile strategies that...
The production of biologics that treat complex diseases, such as cancer, autoimmune, and infectious disease, requires careful monitoring control cell cultures. While bioprocess optimizations have dramatically improved yields, a lack analytical tools has made it challenging to identify accompanying intracellular improvements. Intracellular redox can diminish the growth productivity biologics-producing cells adversely impact product quality profiles yet characterizing is due its highly...