Antibody humanization process converts any nonhuman antibody sequence into humanized antibodies. This can be achieved using different methods of antibody design and engineering. This chapter will primarily focus on antibody design using a homology model followed by framework shuffling of murine to human germline sequence for humanization. Historically, mouse antibodies have been humanized using sequence-based approaches, in which all the murine frameworks are replaced with most homologous human germline sequence or related scaffold. Most often this humanized antibody design, when tested, has a significantly reduced binding or no binding to the cognate antigen. This is due to noncompatibility of mouse CDRs being supported by non-native human framework scaffold. This mismatch between mouse, human structural fold, and elimination of key conformational residues often leads to antibody humanization failures. Recently, there has been advent of homology modelor structure-guided antibody humanization. Instead of humanization based on linear sequence, this approach takes into account the tertiary structure and fold of the mouse antibody. A mouse homology model of the fragment variable is created, and based on sequence alignment with human germline, residues that are different in mouse are replaced with humanized sequence in the model. Energy minimization is applied to this humanized model that also delineates residues which might have steric clashes due to change in the overall tertiary conformation of the humanized antibody. Therefore, a homology model-guided with rational mutations, and reintroduction of key conformational residues from mouse antibody not only eliminates steric clashes but might also restore function in relation to binding affinity to its antigen.
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Exp Neurol
January 2025
CERVO Brain Research Centre, Québec, Québec G1J 2G3, Canada; Department of Psychiatry and Neuroscience, Université Laval, Québec City G1V 0A6, Canada. Electronic address:
Chronic cerebral hypoperfusion induced by permanent unilateral common carotid artery occlusion in mice was recently found to induce an age-dependent formation of insoluble cytoplasmic TDP-43 aggregates reminiscent of pathological changes found in human vascular dementia. In this model, the gradual deregulation of TDP-43 homeostasis in cortical neurons was associated with marked cognitive and motor deficits. To target the TDP-43-mediated toxicity in this model, we generated an adeno-associated virus vector encoding a single-chain antibody against TDP-43, called scFv-E6, designed for pan-neuronal transduction following intravenous administration.
View Article and Find Full Text PDFJ Chromatogr A
December 2024
Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi, India. Electronic address:
Recent advancements in technology, such as the emergence of artificial intelligence (AI) and machine learning (ML), have facilitated the progression of the biopharmaceutical industry toward the implementation of Industry 4.0. As per the guidelines set by the USFDA, process validation for biopharmaceutical production consists of three stages: process design, process qualification, and continuous process verification (CPV).
View Article and Find Full Text PDFPLoS Pathog
January 2025
Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS) and Shanghai Institute of Infectious Disease and Biosecurity, Shanghai Frontiers Science Center of Pathogenic Microorganisms and Infection, School of Basic Medical Sciences, Fudan University, Shanghai, China.
Vaccines are widely regarded as one of the most effective strategies for combating infectious diseases. However, significant challenges remain, such as insufficient antibody levels, limited protection against rapidly evolving variants, and poor immune durability, particularly in subunit vaccines, likely due to their short in vivo exposure. Recent advances in extending the half-life of protein therapeutics have shown promise in improving drug efficacy, yet whether increasing in vivo persistence can enhance the efficacy of subunit vaccines remains underexplored.
View Article and Find Full Text PDFAngew Chem Int Ed Engl
January 2025
University at Buffalo - The State University of New York, Chemistry, 679 NSC, 14260-3000, Buffalo, UNITED STATES OF AMERICA.
Domain antibodies such as monobodies provide an attractive immunoglobin fold for evolving high-affinity protein binders targeting the intracellular proteins implicated in cell signalling. However, it remains a challenge to endow cell permeability to these small and versatile protein binders. Here, we report a streamlined approach combining orthogonal crosslinking afforded by a genetically encoded β-lactam-lysine (BeLaK) and genetic supercharging to generate cell-penetrating monobodies.
View Article and Find Full Text PDFRheumatol Int
January 2025
Department of Rheumatology, Medical University of Lodz, Łódź, Poland.
Scleromyxedema is a rare chronic fibromucinous disorder characterized by a generalized papular and sclerodermoid eruption. Despite its clinical significance, no definitive therapeutic guidelines exist for scleromyxedema, making management challenging. Herein, we present a case of a 76-year-old female patient referred for evaluation of systemic sclerosis, presenting with distinctive cutaneous manifestations and neurological symptoms.
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