Dear Ayesha Fahim . Concerning your issue about the method to extract cell membrane proteins of c. albicans from a mixed species biofilm without using a kit. The opportunistic human fungal pathogen Candida albicans causes a wide variety of infections including deep systemic syndromes. The C. albicans plasma membrane is an important interface in the host-pathogen relationship. The plasma membrane proteins mediate a variety of functions, including sensing and signalling to the external environment, in which the glycosylphosphatidylinositol (GPI)-anchored membrane proteins play a crucial role. A subproteomic approach to obtain a global picture of the protein composition of the C. albicans plasma membrane was developed, and different strategies were tested in order to extract the largest number of yeast plasma membrane proteins and GPI-anchored membrane proteins. These methods involved: (i) protoplast generation, (ii) mechanical disruption, (iii) ultracentrifugation in sucrose gradients, and (iv) Na(2)CO(3) treatments. To isolate GPI-anchored proteins two additional steps were performed: two-phase separation and phosphatidylinositol-phospholipase C treatment. After LC-MS/MS analysis using both a MALDI-TOF/TOF and a linear ion trap quadrupole, a total of 214 membrane proteins were identified, including 41 already described as plasma membrane proteins, 20 plasma membrane associated proteins, and 22 proteins with unknown membrane localisation. Bioinformatic analysis revealed that this set of C. albicans membrane proteins is highly enriched in proteins involved in biopolymer biosynthesis or transport processes. Furthermore, after phosphatidylinositol-phospholipase C treatment, 12 GPI-anchored membrane proteins were released and identified; most of them are associated with cell wall beta-glucan synthesis and maintenance or are virulence factors, such as phospholipases or aspartyl proteinases. Different techniques have been used to extract cell wall components of C. albicans. These include physical, chemical, and enzymatic methods and a combination of them. The choice of extracting reagents and techniques, the sequence of extraction methods, and the use of either intact cells or purified cell walls as the starting material may affect both qualitative and quantitative solubilization of cell wall components. In general, and due to the insolubility of both chitin and glucans, sequential alkali and acid treatments are required to effect their extraction. In early studies, mannans were extracted from whole cells or isolated cell walls by alkali treatment and further precipitated with Fehling’s solution as a copper complex . A milder procedure involved mannan extraction from cells resuspended in citrate buffer (pH 7) by autoclaving and further purification by precipitation with Fehling’s solution or with Cetavlon. This topic is covered more extensively in other reviews. I think the following below links may help you in your analysis:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC98909/

https://www.ncbi.nlm.nih.gov/pubmed/19824013

https://www.ncbi.nlm.nih.gov/pubmed/10768782

Thanks