Salp15, a 15-kDa salivary gland protein plays an important role in tick blood-feeding and transmission of Borrelia burgdorferi, the causative agent of Lyme borreliosis. The comparative studies reveal that Salp15 is a genetically conserved protein across various Ixodes species. In this study, we have identified a Salp15 homolog, designated as Iaff15, from Ixodes affinis ticks that are the principal enzootic vectors of B. burgdorferi sensu stricto in the southeastern part of the United States. Comparison of the annotated amino acid sequences showed that Iaff15 share 81% homology with I. sinensis Salp15 homolog and 64% homology with I. scapularis Salp15. Phylogenetic analysis revealed that Iaff15 come within the same clade with I. sinensis, I. scapularis, and I. pacificus Salp15 homologs. The bioinformatics analysis of the posttranslational modifications prediction revealed that all the Salp15 family members contain glycosylation sites. In addition, Iaff15 carried a higher number of Casein Kinase II phosphorylation sites in comparison to the other Salp15 family members. Collectively, high sequence conservation distributed over the entire amino acids sequence not only suggests an important role for Iaff15 in I. affinis blood feeding and vector-pathogen interactions but may also lead to the development of an anti-vector vaccine against this group of ticks.