Background: Developing world are always looking for monitoring tools during reagent shortage and equipments troubles which are very frequent. The aim of this study was to evaluate Serum Protein Electrophoresis (SPE) as a marker for assessing HIV treatment response. Methods: A cross-sectional study was conducted with 220 participants in four distinct groups: Symptomatic HIV positive patients [specifically those on antiretroviral treatment (ART) versus those not on ART] asymptomatic HIV positive patients, and healthy blood donors. Five serum protein fractions (Albumin, Alpha-1, Alpha-2, Beta, and Gamma) were compared between these groups after measuring the density of the fractions. Results: Concentration of gamma globulin was lowest among healthy blood donors, intermediate and comparable among asymptomatic HIV positive and symptomatic HIV positive on ART and highest among untreated symptomatic HIV positive. Concentration of gamma globulin was inversely correlated with the disease stage (p < 0.001). Conclusion: In this study, conducted in a setting where the burden of infectious diseases is high, the density of gamma globulin and albumin fractions were significantly associated with HIV status, and among HIV positive patients, with stage of HIV disease and ART. These results suggest that the feasibility of using SPE for monitoring the response of ART in low resource settings should be further explored.