Breast cancer is the most frequent kind of invasive cancer in women. Biomarker monitoring and prognosis provide outstanding clinical information that may be employed to battle various cancers. This study aimed to examine a variety of biomarkers utilizing a multiplex bead array. BTLA, CD27, CD28, TIM-3, HVEM, CD40, LAG-3, TLR-2, PD-1, CD80, CD86, PDL-1, PDL-2, and ICOS are common biomarker checkpoint proteins found in a selective panel. This panel examined a research group of 59 patients with BC and 17 healthy people. The expression patterns of a handful of the fourteen biomarkers varied considerably between people with BC and healthy controls. With P-values of (p= 0.05), (p= 0.02), (p= 0.01), and (p= 0.02), general features of the two groups of malignant and non-malignant patients revealed significant correlations in parameters such as the age of first menstruation, pregnancy, menopausal status, and hormone replacement treatment. With P- values, there was a substantial rise in BTLA, HVEM, TLR-2, and PDL-1 serum levels, as well as a significant drop in CD86. Furthermore, these markers indicated a significant relationship with the clinic-pathological aspects of the patients. With a P-value of 0.0002, both BTLA and HVME showed a statistically significant connection with hormone receptor phenotypes. For ER status, BTLA, CD86, and PDL1 had equivalent significance values of (p= 0.0148), (p= 0.0166), and (p= 0.0001). BTLA was related to lymph node involvement (p=0.0397), whereas TLR2 was associated with HER2 status (p=0.0332). BTLA, HVEM, TLR-2 CD86, and PDL-1 may be valuable biomarkers for cancer monitoring and prognosis.