PepLess: A computational tool developed to optimize SPOT synthesis and analyze related immunoassay data

SPOT synthesis is a simple and relatively inexpensive technique for synthesizing a large number of peptides on a cellulose membrane. However, this method becomes exhaustive and impractical when handling a large quantity of sequences with diverse specifications, often resulting in the generation of redundant peptides. Addressing this challenge, we present PepLess, an innovative computational tool designed to analyze multiple sequences simultaneously, thereby reducing the utilization of spot synthesis membranes. By avoiding the production of redundant peptides, PepLess offers a substantial improvement in efficiency and cost-effectiveness, minimizing both expenses and equipment usage. This work outlines the challenges associated with excessive peptide generation, introduces the functionality and benefits of PepLess, and concludes with validation through a case study employing Peruvian-origin Loxosceles laeta spider transcriptome sequences for SPOT membrane production. In this case study we observed approximately a 21 % reduction in required spots for membrane production. Another case study was also performed applying the spike glycoprotein sequences of SARS-CoV-2, and a reduction of 75 %–77 % was identified. The analytical technique within PepLess enables clearer and easier identification of the identified, providing an efficient and automated approach to peptide analysis. • PepLess tool automates SPOT synthesis, reducing redundancy and manual analysis. • Efficiently handles extensive peptide sequences, improving large-scale sequence analysis. • Streamlines peptide synthesis and immunization protocols with optimized methodologies. • Advances the field with computational solutions for managing repetitive protein regions.