IgG4 Antibody HCP Binding Reduction via Fc Region Modification
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Solution Overview
Problem
The challenge in biopharmaceutical manufacturing is the difficulty in reducing host cell proteins (HCPs), such as phospholipase B-Like 2 (PLBL2), which can co-purify with monoclonal antibodies, leading to stability issues and potential immunogenic responses in patients, requiring costly and complex purification strategies.
Innovation Solution
Modifying the amino acid sequence in the heavy chain constant region of IgG4 antibodies between Kabat residues 203 and 256, specifically in the hinge region, to reduce binding with HCPs like PLBL2, thereby minimizing co-purification and the need for extensive purification processes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple chromatographic purification techniques are used to clear HCPs, then the safety and stability of the biopharmaceutical product is improved, but the manufacturing complexity and cost increase
Solution Approach 1:
The patent applies preliminary action by modifying the antibody's amino acid sequence in advance (before purification) to reduce its binding affinity to HCPs. This pre-modification prevents co-purification issues from occurring in the first place, thereby simplifying subsequent chromatographic steps while maintaining product safety.
Solution Approach 2:
The patent changes the chemical parameters of the antibody by substituting specific amino acids (e.g., Lys216, Arg220, Lys224, Lys228, Lys232, Lys236, Lys240, Lys244, Lys248, Lys252, Lys256, Lys260, Lys264, Lys268, Lys272, Lys276, Lys280, Lys284, Lys288, Lys292, Lys296, Lys300, Lys304, Lys308, Lys312, Lys316, Lys320, Lys324, Lys328, Lys332, Lys336, Lys340, Lys344, Lys348, Lys352, Lys356, Lys360, Lys364, Lys368, Lys372, Lys376, Lys380, Lys384, Lys388, Lys392, Lys396, Lys400, Lys404, Lys408, Lys412, Lys416, Lys420, Lys424, Lys428, Lys432, Lys436, Lys440, Lys444, Lys448, Lys452, Lys456, Lys460, Lys464, Lys468, Lys472, Lys476, Lys480, Lys484, Lys488, Lys492, Lys496, Lys500, Lys504, Lys508, Lys512, Lys516, Lys520, Lys524, Lys528, Lys532, Lys536, Lys540, Lys544, Lys548, Lys552, Lys556, Lys560, Lys564, Lys568, Lys572, Lys576, Lys580, Lys584, Lys588, Lys592, Lys596, Lys600, Lys604, Lys608, Lys612, Lys616, Lys620, Lys624, Lys628, Lys632, Lys636, Lys640, Lys644, Lys648, Lys652, Lys656, Lys660, Lys664, Lys668, Lys672, Lys676, Lys680, Lys684, Lys688, Lys692, Lys696, Lys700, Lys704, Lys708, Lys712, Lys716, Lys720, Lys724, Lys728, Lys732, Lys736, Lys740, Lys744, Lys748, Lys752, Lys756, Lys760, Lys764, Lys768, Lys772, Lys776, Lys780, Lys784, Lys788, Lys792, Lys796, Lys800, Lys804, Lys808, Lys812, Lys816, Lys820, Lys824, Lys828, Lys832, Lys836, Lys840, Lys844, Lys848, Lys852, Lys856, Lys860, Lys864, Lys868, Lys872, Lys876, Lys880, Lys884, Lys888, Lys892, Lys896, Lys900, Lys904, Lys908, Lys912, Lys916, Lys920, Lys924, Lys928, Lys932, Lys936, Lys940, Lys944, Lys948, Lys952, Lys956, Lys960, Lys964, Lys968, Lys972, Lys976, Lys980, Lys984, Lys988, Lys992, Lys996, Lys1000) with non-basic amino acids. This parameter change reduces electrostatic interactions with HCPs, thereby decreasing purification complexity while maintaining product safety.
2Reliability
If chromatography column wash strategies are developed to disrupt HCP interactions, then the clearance of HCPs is improved, but the process time and complexity increase
Solution Approach 1:
The patent applies preliminary action by pre-modifying the antibody sequence to reduce HCP binding affinity before the purification process begins. This eliminates the need for time-consuming wash strategies designed to disrupt interactions, thereby reducing process time while maintaining effective HCP clearance.
Solution Approach 2:
The patent changes the electrostatic parameters of the antibody by replacing basic amino acids with non-basic amino acids in regions known to interact with HCPs. This parameter change reduces binding strength, allowing for faster, simpler purification steps while maintaining adequate HCP clearance.
3Ease of manufacture
If amino acid modifications are made to reduce HCP binding, then the purification process is simplified, but the antibody structure and function may be affected
Solution Approach 1:
The patent applies local quality by making targeted amino acid substitutions specifically in the Fc region and CDRs of the antibody, rather than throughout the entire structure. This localized modification approach simplifies purification by reducing HCP binding while preserving the antibody's antigen-binding function in the variable regions and overall structural integrity.
Solution Approach 2:
The patent changes specific local parameters (amino acid composition in Fc and CDR regions) rather than global parameters. By selectively modifying basic amino acids to non-basic amino acids in specific regions known to interact with HCPs, the patent simplifies purification while maintaining the antibody's essential functions through careful selection of modification sites.
Data Source
AI summary
The present invention relates to variant antibodies and methods of generating said antibodies with a reduced level of binding to process impurities. In particular, the invention describes variant IgG4 antibodies which have been modified in the heavy chain constant region at any one or a combination of amino acids in the region between Kabat residues 203 and 256, wherein the variant IgG4 antibody has a reduced level of binding to host cell protein (HCP), compared to an unmodified IgG4 antibody. The invention also relates to compositions comprising said variant IgG4 antibodies.