Chiral HPLC stationary phases based on immobilized biopolymers (i.e., proteins, enzymes, antibodies etc.) are of growing importance for the separation and purification of enantiomeric compounds, especially in the areas of biological, pharmaceutical, and clinical chemistry. In the work described in this paper, several silica-immobilized serum albumin phases were synthesized and characterized by a variety of techniques. Low-temperature nitrogen adsorption isotherms were measured and employed to evaluate the surface characteristics of the stationary phases as a function of both the immobilization matrix and the surface modification chemistry. Elemental analysis and high resolution thermogravimetry were used to evaluate the surface coverages at each stage of modification. Based on these results, surface parameters such as the specific surface area, pore size distribution, bonding density, etc. were calculated, as well as the changes of these parameters as a function of the modification chemistry were analyzed. Different silicas of varying mean pore diameter from 30 nm to 400 nm were used.