The urgent need for gluten-free bread by Celiac and gluten-sensitive populations has spawned a new and emerging food industry to provide substitutes for gluten-free bread. This study aims to provide an effective substitute bread using locally available indigenous crops like Jackfruit Seed and Flour (Artocarpus heterophyllus Lam) and Mapillai Samba Rice Flour (Bridegroom Oryza sativa L) as the key ingredients. Central Composite Rotatable Design (CCRD) of Response Surface Methodology (RSM) was applied to study the overall comparative acceptability of the gluten-free bread. Gluten-free bread was prepared using 20 variations of composite flour without gluten by varying the proportions of these three major ingredients. The best combination of the three major ingredients for the production of gluten-free bread was determined by studying the textural properties (hardness and springiness), loaf volume, colour value and sensory quality. Such a viable and desirable product will have commercial implications.