Rationale. Direct identification of peptides presented within major histocompatibility complexes class I (HLA I) on the surface of tumor cells can be used to develop targeted immunotherapy for ovarian adenocarcinoma. However, standard immunopeptidomic analysis protocols can vary significantly in the efficiency of immunopeptidome isolation depending on lysis conditions. Optimization of detergents for immunoaffinity isolation of complexes allows for increased assay sensitivity, expansion of the repertoire of identifiable ligands, and an increased likelihood of detecting clinically significant peptides suitable for the development of personalized immunotherapeutic approaches. Objective. To evaluate the effect of various detergents on the efficiency of HLA complex peptide ligand identification. Methods. Immunopeptidomes from cell lines were identified using immunoaffinity chromatography followed by chromatograph mass spectrometry analysis. Results. In this study, we tested an immunoaffinity chromatograph protocol for immunopeptide isolation, comparing various detergents (CHAPS, NP-40, SOD, and Triton X-100) for cell lysis. No statistically significant differences in the number of identified peptides were observed. Conclusion. Using each of the four tested detergents resulted in the identification of unique sets of peptides.