Multiplex determination of serological signatures in the sera of colorectal cancer patients using hydrogel biochips

Abstract

Colorectal cancer (CRC) is the third most common malignancy in industrialized countries. Despite the advances in diagnostics and development of new drugs, the 5‐year survival remains only 60–65%. Our approach to early diagnostics of CRC is based on the determination of serological signatures with an array of hemispherical hydrogel cells containing immobilized proteins and oligosaccharides (glycochip). The compounds immobilized on the glycochip include tumor‐associated glycans (SiaTn, Tn, TF, LeC, LeY, SiaLeA, and Manβ1‐4GlcNAcβ) and antibodies against human immunoglobulins IgG, IgA, and IgM. The glycochip detects antibodies against tumor‐associated glycans in patients’ sera. The simultaneous measurement of the levels of immunoglobulins enhances the diagnostic impact of the signatures. In this work, we found previously unreported increase in antibodies against oligosaccharide Manβ1‐4GlcNAcβ in patients with CRC. In parallel with these experiments, we determined the levels of oncomarkers carcinoembryonic antigen (CEA), cancer antigen (CA) 19–9, CA 125, CA 15–3, human chorionic gonadotropin (HCG), and alpha‐fetoprotein (AFP) using another gel‐based biochip with immobilized antibodies (oncochip) developed earlier in our laboratory. In total, 69 samples from healthy donors, 33 from patients with colorectal carcinoma, and 27 from patients with inflammatory bowel diseases were studied. The use of combined signatures of antiglycan antibodies and oncomarkers provides much better predictive value than the conventional measurement of oncomarkers CEA and CA 19–9. Positive predictive value of CRC diagnoses using together glycochip and oncochip reached 95% with the sensitivity and specificity 88% and 98%, respectively. Thus, the combination of antibody profiling with detection of conventional oncomarkers proved to be a promising tool in diagnostics of CRC.

Human Natural Antibodies Recognizing Glycan Galβ1-3GlcNAc (LeC)

“…Third, these antibodies bind B cells in tumor lesion milieu [5]. Fourth, in studies aimed at finding diagnostic signatures (a signature usually consists of 6-10 anti-glycan nAbs), these antibodies turned out to be the most frequent constituent of the signature [6][7][8]. In addition, two monoclonal antibodies with similar specificities are known-LU-BCRU-G7, which specifically binds to breast cancer tissue [9] and 58-1, which was generated using CA19.9 glycoprotein as an immunogen [10] (Specificity and comparison of monoclonal antibodies (mAb) with human anti-Le C are presented in Reference [10]).…”

Blood-Based Protein Signatures for Early Detection of Colorectal Cancer: A Systematic Review

“…The majority of studies were conducted in Europe (21/36). Of these, four studies were carried out in Denmark, 21 , 22 , 23 , 24 three each in Germany, 25 , 26 , 27 Spain, 28 , 29 , 30 and Russia, 31 , 32 , 33 two each in Finland 34 , 35 and the United Kingdom, 36 , 37 one study each in Italy, 38 France, 39 Poland, 40 and Czech Republic, 26 and at last one study was multicenter. 41 Eleven studies that were carried out in Asia, included eight studies from China, 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 two studies from Japan, 50 , 51 and one study from Korea.…”

Signatures of Anti-Thomsen — Friedenreich Antigen Antibody Diversity in Colon Cancer Patients

“…Alternatively, it may be speculated that higher sialylated TF-specific antibodies would The currently dominating approach in anti-glycan antibody studies on cancer patients is the antibody levels profiling. The analysis employs multi-glycan microarray techniques [35,55,56] to find cancerspecific changes based on antibody profiling and/ or multiplex combination of more informative antibodies. However, given the extremely high structural and functional polymorphism of anti-glycan antibodies, we believe that a more promising approach might be a search for disease-specific antibody subsets.…”