The field of biotherapeutics has seen rapid advancements with antibodies and antibody-like formats emerging as a major class of therapeutic proteins. For scientists and researchers in biotech, understanding the structural and dynamic characteristics of antibodies is crucial. These insights inform the engineering of more effective therapeutic antibodies and vaccines.
Antibodies are Y-shaped molecules composed of two identical heavy chains and two identical light chains. This classic IgG architecture is modular, allowing for various engineering modifications to enhance biophysical properties and therapeutic potential. The antibody’s “stem” forms the crystallizable fragment (Fc), involved in interactions with cell surface receptors, while the “arms” or antigen-binding fragments (Fab) contain variable domains responsible for antigen recognition. An important part of antibody structure is the immunoglobulin fold (Ig-fold), a conserved feature across various proteins. Each Ig-like domain consists of two anti-parallel β-sheets stabilized by disulfide bridges. The variable domains (VH and VL) and constant domains (CH1-CL and CH3-CH3) form heterodimers and homodimers, respectively, influencing the antibody’s functional properties.
The Role of Dynamics in Antibody Function
Antibodies are not static structures; their dynamic nature is central to their function. Molecular dynamics simulations reveal that antibodies exist as conformational ensembles, with different rearrangements occurring on various timescales. Subtle movements, such as bond vibrations and side-chain fluctuations, occur on the nanosecond scale, while larger loop rearrangements take milliseconds or longer. This dynamic behavior allows antibodies to adopt multiple conformations, enhancing their ability to recognize diverse antigens.
Predicting antibody structures from amino acid sequences remains a challenge, particularly for the hypervariable complementarity determining region (CDR) loops. Traditional template-based approaches, like homology modeling, have limitations due to structural diversity. However, hybrid methods and artificial intelligence-driven models, such as AlphaFold2 and DeepAB, show promise in improving accuracy, especially for CDR loops. Incorporating information on the dynamic nature of antibodies, through techniques like NMR spectroscopy and cryo-EM, can further enhance prediction models.
Innovations and Experimental Techniques in Antibody Engineering
High-resolution structural techniques, such as X-ray crystallography, NMR spectroscopy, and cryo-EM, are instrumental in understanding antibody-antigen recognition. Each method offers unique advantages. X-ray crystallography provides atomic-resolution structures, NMR spectroscopy captures dynamics in solution, and cryo-EM facilitates the study of large complexes in near-physiological conditions. Recent advancements in cryo-EM, particularly the resolution revolution, have made it a preferred method for detailed epitope mapping. This technique can process heterogeneous samples, enabling comprehensive analysis of polyclonal antibody responses and guiding the design of novel therapeutics.
Antibody engineering has led to the development of various formats to meet diverse therapeutic needs. Bispecific antibodies, for example, can simultaneously recognize two different epitopes, enhancing their therapeutic potential. Engineering techniques ensure correct heavy and light chain pairing, optimizing heterodimerization for desired functionalities. Smaller antibody formats, such as single-chain variable fragments (scFv) and diabodies, offer advantages in expression and pharmacokinetics. These formats retain the binding specificity of conventional antibodies but with improved properties for therapeutic applications.
References:
- L., M., Pomarici, N. D., Fischer, A., Hoerschinger, V. J., Kroell, K. B., Riccabona, J. R., Kamenik, A. S., Loeffler, J. R., Ferguson, J. A., Perrett, H. R., Liedl, K. R., Han, J., & Ward, A. B. (2023). Structure and Dynamics Guiding Design of Antibody Therapeutics and Vaccines. Antibodies, 12(4), 67. https://doi.org/10.3390/antib12040067