A wide diversity of serological biomarkers is available for clinic diagnosis but individually their lack of specificity decreases their diagnostic and prognostic values. Thus, there is an urgent need to develop more sensitive, reliable and specific screening tools for biomarkers detection. Protein microarrays have demonstrated to be powerful for screening and identifying biomarkers from minute samples. However, efficient multiplex analysis remains challenging due to biomarkers variability. A lot of parameters (surface chemistry, composition and pH of the spotting buffer, blocking reagents, antibody concentration, etc.) have to be optimized for obtaining sensitive and specific detection. We have previously developed and characterized various surface chemistries allowing covalent (NHS ester activated surface, activated CMD surface, MAMVE surface) or non covalent immobilization (carboxylic surface, chitosan surface, aminated surface) of proteins. This work is focused on the development of customized antibody and antigen microarrays implemented with various surface chemistries, for the sensitive detection of biomarkers in cancer sera.