Improvement of Biophysical Properties and Affinity of a Human Anti-L1CAM Therapeutic Antibody through Antibody Engineering Based on Computational Methods.

The biophysical properties of therapeutic antibodies influence their manufacturability, efficacy, and safety. To develop an anti-cancer antibody, we previously generated a human monoclonal antibody (Ab417) that specifically binds to L1 cell adhesion molecule with a high affinity, and we validated its anti-tumor activity and mechanism of action in human cholangiocarcinoma xenograft models. In the present study, we aimed to improve the biophysical properties of Ab417. We designed 20 variants of Ab417 with reduced aggregation propensity, less potential post-translational modification (PTM) motifs, and the lowest predicted immunogenicity using computational methods. Next, we constructed these variants to analyze their expression levels and antigen-binding activities. One variant (Ab612)-which contains six substitutions for reduced surface hydrophobicity, removal of PTM, and change to the germline residue-exhibited an increased expression level and antigen-binding activity compared to Ab417. In further studies, compared to Ab417, Ab612 showed improved biophysical properties, including reduced aggregation propensity, increased stability, higher purification yield, lower pI, higher affinity, and greater in vivo anti-tumor efficacy. Additionally, we generated a highly productive and stable research cell bank (RCB) and scaled up the production process to 50 L, yielding 6.6 g/L of Ab612. The RCB will be used for preclinical development of Ab612.

The medicinal plant Porana volubilis contains polysaccharides with anticoagulant activity mediated by heparin cofactor II.

We searched for polysaccharides with anticoagulant activity and inhibitory action on platelet aggregation induced by collagen in 59 species of medicinal plants. We then concentrated our studies on the polysaccharide from the species Porana volubilis, which showed the highest anticoagulant activity among the plants tested. The polysaccharide from this species has an average molecular mass of approximately 10 kDa, contains mainly galactose, galacturonic acid, and mannose but no sulfate esters. Its anticoagulant activity is mediated by the enhancement of thrombin inhibition that in turn is mediated by heparin cofactor II but not by antithrombin. The galacturonic acid residues are essential for activity since after reduction of its carboxyl groups the anticoagulant activity disappears. Our report is the first description of a natural nonsulfated polysaccharide from higher plants with anticoagulant activity, which may constitute a new source of compounds with action on coagulation and, perhaps, on thrombosis.