The Effect of Physicochemical Modification on the Function of Antibodies Induced by Anti-Nicotine Vaccine in Mice

Abstract

Smoking remains one of the major causes of morbidity and mortality worldwide. One approach to assisting smoking cessation is via anti-nicotine vaccines, composed of nicotine-like haptens conjugated to a carrier protein plus adjuvant(s). We have previously shown that the carrier, hapten, linker, hapten load, degree of conjugate aggregation, and presence of adducts can each influence the function (nicotine-binding capacity) of the antibody (Ab) induced. Herein, we extend those findings and show that tertiary structure is also critical to the induction of functional immune responses and that this can be influenced by conjugation conditions. We evaluated immunogenicity in mice using six lots of NIC7-CRM, a conjugate of 5-aminoethoxy-nicotine (Hapten 7), and a single point (glycine 52 to glutamic acid) mutant nontoxic form of diphtheria toxin, cross-reactive material 197 (CRM197), which were synthesized under different reaction conditions resulting in conjugates with equivalent molecular characteristics (hapten load, aggregates, adducts), but a different tertiary structure. When tested in mice, better functional responses (reduced nicotine in the brain of immunized animals relative to non-immunized controls) were obtained with conjugates with a more closed structure than those with an open conformation. These studies highlight the need for a better understanding of the physicochemical properties of small molecule conjugate vaccines.

Preclinical Efficacy and Characterization of Candidate Vaccines for Treatment of Opioid Use Disorders Using Clinically Viable Carrier Proteins

“…46 Instead, drug-conjugate vaccines containing smaller carrier proteins, such as BSA or OVA 39 and TT or CRM 197, 18 can be characterized by MALDI-TOF or other analytical assays. 47 In fact, conjugation conditions are often optimized using BSA and OVA in lieu of larger carrier proteins because it is easier to measure their haptenation ratio. 40,48–50…”

“…Previous studies tested this hypothesis by immunizing subjects with several batches of nicotine vaccines displaying various degrees of aggregation, and found that the extent of aggregation (and other physical chemical properties of the conjugates) correlated with individual vaccine efficacy against nicotine in both mice and nonhuman primates. 69–71…”

Pengaruh injeksi nikotin terhadap motilitas, viabilitas, dan integritas membran spermatozoa mencit (Mus Musculus)

“…Proses pembentukan spermatozoa sangat sensitif terhadap pengaruh dari luar misalnya yang ditimbulkan oleh asap rokok. Nikotin memiliki partikel yang sangat kecil untuk mengaktifkan sistem imun dan dapat melewati sawar darah otak (Thorn et al, 2017).…”

Smoking Cessation Treatments: Current Psychological and Pharmacological Options

“…It would be indicated for the following groups of subjects: (a) heavy smokers who have made multiple unsuccessful attempts to quit using different methods; (b) patients who are highly motivated to quit; and (c) patients who have positive attitudes toward vaccines 20 . Modifications designed to improve serum antibody affinity are being tested and may improve the performance of future clinical candidate vaccines [21][22][23] .…”