Epitope binning for multiple antibodies simultaneously using mammalian cell display and DNA sequencing.

Epitope binning, an approach for grouping antibodies based on epitope similarities, is a critical step in antibody drug discovery. However, conventional methods are complex, involving individual antibody production. Here, we established Epitope Binning-seq, an epitope binning platform for simultaneously analyzing multiple antibodies. In this system, epitope similarity between the query antibodies (qAbs) displayed on antigen-expressing cells and a fluorescently labeled reference antibody (rAb) targeting a desired epitope is analyzed by flow cytometry. The qAbs with epitope similar to the rAb can be identified by next-generation sequencing analysis of fluorescence-negative cells. Sensitivity and reliability of this system are confirmed using rAbs, pertuzumab and trastuzumab, which target human epidermal growth factor receptor 2. Epitope Binning-seq enables simultaneous epitope evaluation of 14 qAbs at various abundances in libraries, grouping them into respective epitope bins. This versatile platform is applicable to diverse antibodies and antigens, potentially expediting the identification of clinically useful antibodies.

Are the Clinical Results of Locking Plate Fixation for Distal Radius Fractures Inferior in Patients over 80 Years of Age? A Multicentre (TRON Group) Study.

Introduction: With the aging of the population, the proportion of distal radius fracture patients who are > 80 years of age is increasing. In this study, we compared the postoperative clinical and radiographic outcomes between super-elderly patients (age: ≥ 80 years) and middle-elderly (age: 65-79 years) who were treated with volar locking plate (VLP) fixation for distal radius fractures. Patients and Methods: Patients of > 65 years of age with distal radius fractures treated by VLP fixation between 2015 and 2019, and who were followed for at least 6 months after surgery were included in our database (named TRON). Patients with open fractures, multiple-trauma, or who received fixation with implants other than a VLP were excluded. We evaluated postoperative complications, Mayo wrist score (MWS), and radiographic outcomes. Results: We identified 589 patients in this study; 452 were 65-79 years of age (Group A) and 137 were ≥ 80 years of age (Group B). After propensity score matching, we evaluated 309 patients in Group A and 103 patients in Group B. The mean follow-up period was 10.7 ± 4.6 months. Twenty-eight patients (9.1%) in Group A and 5 patients in Group B (4.9%) experienced post-operative complications (non-significant: p = 0.212). The postoperative MWS at 1, 3, and 6 months, respectively, was 65.4 ± 11.7, 75.2 ± 11.0, and 79.6 ± 10.5 in Group A and 67.1 ± 9.61, 75.7 ± 10.7, and 80.6 ± 9.7 in Group B (non-significant: p = 0.418, 0.893, 0.452, respectively). The differences in volar tilt, radial inclination, ulnar variance between the postoperative and last follow-up radiographs did not differ between the two groups to a statistically significant extent (p = 0.053, 0.437, 0.529, respectively). Conclusion: Our study showed that the clinical and radiographic outcomes of distal radius fractures treated with VLP in super-elderly patients were comparable to those in middle-elderly patients.

Synthesis of enantiomerically enriched drug precursors and an insect pheromone via reduction of ketones using commercially available carbonyl reductase screening kit "Chiralscreen® OH".

Commercially available "Chiralscreen® OH" starter kit containing five types of carbonyl reductases (E001, E007, E031, E039, and E078) was used for the reduction of several aromatic and aliphatic ketones to obtain enantiomerically enriched drug precursors and an insect pheromone. Almost stereochemically pure secondary alcohols, used in the synthesis of drugs such as (R)-rasagiline mesylate, (S)-rivastigmine, (R)-chlorphenesin carbamate, and (R)-mexiletine, and the insect pheromone (4S,5R)-sitophilure, were conveniently obtained. The enzymes worked well with ketones containing at least one non-bulky substituent at the carbonyl group. The diverse stereochemical preference of the above five carbonyl reductases was clarified.

Enoxacin with UVA Irradiation Induces Apoptosis in the AsPC1 Human Pancreatic Cancer Cell Line Through ROS Generation.

Pancreatic cancer is one of the deadliest human cancers. In the current study, we investigated the possibility of a new treatment strategy using a combination of the new fluoroquinolone, enoxacin, and mild ultraviolet A (UVA) irradiation. Enoxacin with UVA irradiation increased the number of annexin V-positive (apoptotic) pancreatic cancer cells in time- and concentration-dependent manners, whereas alone neither had these effects. In addition, enoxacin with UVA irradiation induced cleavage of poly (ADP-ribose) polymerase in AsPC1 human pancreatic cancer cells. Moreover, the singlet oxygen scavengers, histidine and sodium azide, and the hydroxyl radical scavenger, mannitol, significantly suppressed apoptosis induced by enoxacin and UVA irradiation, respectively. These results suggest that UVA irradiation activates enoxacin, after which activated enoxacin induces apoptosis of AsPC1 cells through generation of reactive oxygen species. Therefore, the combination of enoxacin with mild UVA irradiation may be a useful method for treating pancreatic cancer.