An antidote approach to reduce risk and broaden utility of antibody-based therapeutics

Portnoff, Alyse D.; Gao, Cuihua; Borrok, M. Jack; Gao, Xizhe; Gao, Changshou; Rainey, G. Jonah

doi:10.1074/jbc.m117.775528

Cited by 2 publications

(2 citation statements)

References 42 publications

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“…Thus, these substitutions might not change the antibody's affinity for FcγRIIIa of IgG2_YLA. In relation to FcRn binding, it has been reported that residue IgG1 L309 is included in the key contact positions on the Fc region with FcRn . However, compared to that of IgG2, IgG2_YLA has almost the same KD with FcRn.…”

Section: Discussionmentioning

confidence: 99%

Engineering a human IgG2 antibody stable at low pH

et al. 2020

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IgG2 subclass antibodies have unique properties that include low effector function and a rigid hinge region. Although some IgG2 subclasses have been clinically tested and approved for therapeutic use, they have a higher propensity than IgG1 for aggregation, which can curtail or abolish their biological activity and enhance their immunogenicity. In this regard, acid-induced aggregation of monoclonal antibodies during purification and virus inactivation must be prevented. In the present study, we replaced the constant domain of IgG2 with that of IgG1, using anti-2,4-dinitrophenol (DNP) IgG2 as a model antibody, and investigated whether that would confer greater stability. While the anti-DNP IgG2 antibody showed significant aggregation at low pH, this was reduced for the IgG2 antibody containing the IgG1 CH2 domain. Substituting three amino acids within the CH2 domain-namely, F300Y, V309L, and T339A (IgG2_YLA)-reduced aggregation at low pH and increased CH2 transition temperature, as determined by differential scanning calorimetric analysis. IgG2_YLA exhibited similar antigen-binding capacity to IgG2, low affinity for FcγRIIIa, and low binding ability to C1q. The same YLA substitution also reduced the aggregation of panitumumab, another IgG2 antibody, at low pH. Our engineered human IgG2 antibody showed reduced aggregation during bioprocessing and provides a basis for designing improved IgG2 antibodies for therapeutic applications. K E Y W O R D Sacid-induced aggregate, ADCC, aggregation, antibody, CDC, IgG2 subclass, subclass change

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Section: Discussionmentioning

confidence: 99%

Engineering a human IgG2 antibody stable at low pH

et al. 2020

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“…Although some of these approaches, mostly biotin–streptavidin pair, have been tested in nuclear imaging and therapy in patients, there is still a substantial risks of immunogenicity, , as well as suboptimal clearing efficiency. More recently, several interesting clearing approaches to block FcRn recycling by in vivo PEGylation of the Fc portion or by anti-FcRn antibody have been reported. However, these approaches still retain IgG in tissues, require introduction of non-natural amino acids into the antibody sequence, or interfere with metabolism of natural immunoglobulins.…”

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confidence: 99%

Accelerated Blood Clearance of Antibodies by Nanosized Click Antidotes

et al. 2018

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Long blood half-life is one of the advantages of antibodies over small molecule drugs. At the same time, prolonged half-life is a problem for imaging applications or in the case of antibody-induced toxicities. There is a substantial need for antidotes that can quickly clear antibodies from systemic circulation and peripheral tissues. Engineered nanoparticles exhibit intrinsic affinity for clearance organs (mainly liver and spleen). trans-Cyclooctene (TCO) and methyltetrazine (MTZ) are versatile copper-free click chemistry components that are extensively being used for in vivo bioorthogonal couplings. To test the ability of nanoparticles to eliminate antibodies, we prepared a set of click-modified, clinically relevant antidotes based on several classes of drug carriers: phospholipid-PEG micelles, bovine serum albumin (BSA), and cross-linked dextran iron oxide (CLIO) nanoparticles. Mice were injected with IRDye 800CW-labeled, click-modified IgG followed by a click-modified antidote or PBS (control), and the levels of the IgG were monitored up to 72 h postinjection. Long-circulating lipid micelles produced a spike in IgG levels at 1 h, decreased IgG levels at 24 h, and did not decrease the area under the curve (AUC) and IgG accumulation in main organs. Long-circulating BSA decreased IgG levels at 1 and 24 h, decreased the AUC, but did not significantly decrease organ accumulation. Long-circulating CLIO nanoworms increased IgG levels at 1 h, decreased IgG levels at 24 h, did not decrease the AUC, and did not decrease the organ accumulation. On the other hand, short-circulating CLIO nanoparticles decreased IgG levels at 1 and 24 h, significantly decreasing the AUC and accumulation in the main organs. Multiple doses of CLIO and BSA were not able to completely eliminate the antibody from blood, despite the click reactivity of the residual IgG, likely due to exchange of IgG between blood and tissue compartments. Pharmacokinetic modeling suggests that short antidote half-life and fast click reaction rate should result in higher IgG depletion efficiency. Short-circulating click-modified nanocarriers are the most effective antidotes for elimination of antibodies from blood. This study sets a stage for future development of antidotes based on nanomedicine.

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An antidote approach to reduce risk and broaden utility of antibody-based therapeutics

Cited by 2 publications

References 42 publications

Engineering a human IgG2 antibody stable at low pH

Engineering a human IgG2 antibody stable at low pH

Accelerated Blood Clearance of Antibodies by Nanosized Click Antidotes

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