This collection of review articles features expert content published by RayBiotech Scientists to help accelerate your biomarker discovery efforts.


Cytokines in Cancer Drug Resistance: Cues to New Therapeutic Strategies

Valerie Sloane Jones a,1, Ren-Yu Huang b,1, Li-PaiChen c, Zhe-Sheng Chen d Liwu Fu e, Ruo-PanHuang a, b, f

Abstract – Click here to read the full article

The development of oncoprotein-targeted anticancer drugs is an invaluable weapon in the war against cancer. However, cancers do not give up without a fight. They may develop multiple mechanisms of drug resistance, including apoptosis inhibition, drug expulsion, and increased proliferation that reduce the effectiveness of the drug. The collective work of researchers has highlighted the role of cytokines in the mechanisms of cancer drug resistance, as well as in cancer cell progression. Furthermore, recent studies have described how specific cytokines secreted by cancer stromal cells confer resistance to chemotherapeutic treatments. In order to gain a better understanding of mechanism of cancer drug resistance and a prediction of treatment outcome, it is imperative that correlations are established between global cytokine profiles and cancer drug resistance. Here we discuss the recent discoveries in this field of research and discuss their implications for the future development of effective anti-cancer medicines.

  1. Raybiotech, Inc., 3607 Parkway Lane, Norcross, GA, USA
  2. RayBiotech, Inc., Guangzhou, No. 79 Ruihe Road, Huangpu District, Guangzhou, China
  3. Department of Gynaecologic Oncology, Cancer Institute and Hospital, Guangzhou Medical University, Guangzhou, China
  4. Department of Pharmaceutical Sciences, College of Pharmacy and Health Science, St. John’s University, Queens, NY, USA
  5. Sun Yat-sen University Cancer Center, Guangzhou, China
  6. South China Biochip Research Center, No. 79 Ruihe Road, Huangpu District, Guangzhou, China

Tumor-Induced Perturbations of Cytokines and Immune Cell Networks

Brett Burkholder a, Ren-Yu Huang b, Rob Burgess a, Shuhong Luo a,b, Valerie Sloane Jones a, Wenji Zhang b, Zhi-Qiang Lv b, Chang-Yu Gao b, Bao-Ling Wang b, Yu-Ming Zhang b, Ruo-Pan Huang a,b,c

Abstract – Click here to read the full article

Until recently, the intrinsically high level of cross-talk between immune cells, the complexity of immune cell development, and the pleiotropic nature of cytokine signaling have hampered progress in understanding the mechanisms of immunosuppression by which tumor cells circumvent native and adaptive immune responses. One technology that has helped to shed light on this complex signaling network is the cytokine antibody array, which facilitates simultaneous screening of dozens to hundreds of secreted signal proteins in complex biological samples. The combined applications of traditional methods of molecular and cell biology with the high-content, high-throughput screening capabilities of cytokine antibody arrays and other multiplexed immunoassays have revealed a complex mechanism that involves multiple cytokine signals contributed not just by tumor cells but by stromal cells and a wide spectrum of immune cell types. This review will summarize the interactions among cancerous and immune cell types, as well as the key cytokine signals that are required for tumors to survive immunoediting in a dormant state or to grow and spread by escaping it. Additionally, it will present examples of how probing secreted cell-cell signal networks in the tumor microenvironment (TME) with cytokine screens have contributed to our current understanding of these processes and discuss the implications of this understanding to antitumor therapies.

  1. RayBiotech, Inc., 3607 Parkway Lane, Suite 100, Norcross, GA 30092, USA
  2. RayBiotech, Inc., Guangzhou 510600, China
  3. South China Biochip Research Center, Guangzhou 510630, China

Deciphering Asthma Biomarkers with Protein Profiling Technology

Zhizhou Kuang,1,2,3 Jarad J. Wilson,2 Shuhong Luo,1,2,3 Si-Wei Zhu,1 and Ruo-Pan Huang 1,2,3

Abstract – Click here to read the full article

Asthma is a chronic inflammatory disease of the airways, resulting in bronchial hyperresponsiveness with every allergen exposure. It is now clear that asthma is not a single disease, but rather a multifaceted syndrome that results from a variety of biologic mechanisms. Asthma is further problematic given that the disease consists of many variants, each with its own etiologic and pathophysiologic factors, including different cellular responses and inflammatory phenotypes. These facets make the rapid and accurate diagnosis (not to mention treatments) of asthma extremely difficult. Protein biomarkers can serve as powerful detection tools in both clinical and basic research applications. Recent endeavors from biomedical researchers have developed technical platforms, such as cytokine antibody arrays, that have been employed and used to further the global analysis of asthma biomarker studies. In this review, we discuss potential asthma biomarkers involved in the pathophysiologic process and eventual pathogenesis of asthma, how these biomarkers are being utilized, and how further testing methods might help improve the diagnosis and treatment strain that current asthma patients suffer.

1RayBiotech, Inc., Guangzhou 510600, China
2RayBiotech, Inc., 3607 Parkway Lane, Norcross, GA 30092, USA
3South China Biochip Research Center, Guangzhou 510600, China


Application of multiplex immunoassay technology to investigations of ocular disease

Valerie Sloane Jones (a1), Jian Wu (a2), Si-Wei Zhu (a3) and Ruo-Pan Huang (a1) (a3) (a4)

Abstract – Click here to read the full article

Eye-derived fluids, including tears, aqueous humour and vitreous humour often contain molecular signatures of ocular disease states. These signatures can be composed of cytokines, chemokines, growth factors, proteases and soluble receptors. However, the small quantities (<10 µl) of these fluids severely limit the detection of these proteins by traditional enzyme-linked immunosorbent assay or Western blot. To maximise the amount of information generated from the analysis of these specimens, many researchers have employed multiplex immunoassay technologies for profiling the expression or modification of multiple proteins from minute sample volumes.

(a1)  Raybiotech, Inc., Norcross, GA, USA
(a2)  The Affiliated third Hospital of Sun Yat-sen University, Sun Yat-sen University, Guangdong, China
(a3)  RayBiotech, Inc., Guangzhou, China
(a4) South China Biochip Research Center, Guangzhou, China


Quantitative Screening of Serum Protein Biomarkers by Reverse Phase Protein Arrays

Zhizhou Kuang1,2, Ruochun Huang1,2, Zhimin Yang3,4, Zhiqiang Lv1, Xinyan Chen3,4, Fuping Xu3,4, Yu-Hua Yi1,6, Jian Wu5and Ruo-Pan Huang1,2,6

Abstract – Click here to read the full article

Screening biomarkers in serum samples for different diseases has always been of great interest because it presents an early, reliable, and, most importantly, noninvasive means of diagnosis and prognosis. Reverse phase protein arrays (RPPAs) are a high-throughput platform that can measure single or limited sets of proteins from thousands of patients’ samples in parallel. They have been widely used for detection of signaling molecules involved in diseases, especially cancers, and related regulation pathways in cell lysates. However, this approach has been difficult to adapt to serum samples. Previously, we developed a sensitive method called the enhanced protein array to quantitatively measure serum protein levels from large numbers of patient samples. Here, we further refine the technology on several fronts: 1. simplifying the experimental procedure; 2. optimizing multiple parameters to make the assay more robust, including the support matrix, signal reporting method, background control, and antibody validation; and 3. establishing a method for more accurate quantification. Using this technology, we quantitatively measured the expression levels of 10 proteins: alpha-fetoprotein (AFP), beta 2 microglobulin (B2M), Carcinoma Antigen 15-3(CA15-3), Carcinoembryonic antigen (CEA), golgi protein 73 (GP73), Growth differentiation factor 15 (GDF15), Human Epididymis Protein 4 (HE4), Insulin Like Growth Factor Binding Protein 2 (IGFBP2), osteopontin (OPN) and Beta-type platelet-derived growth factor receptor (PDGFRB) from serum samples of 132 hepatocellular carcinoma (HCC) patients and 78 healthy volunteers. We found that 6 protein expression levels are significantly increased in HCC patients. Statistical and bioinformatical analysis has revealed decent accuracy rates of individual proteins, ranging from 0.617 (B2M) to 0.908 (AFP) as diagnostic biomarkers to distinguish HCC from healthy controls. The combination of these 6 proteins as a specific HCC signature yielded a higher accuracy of 0.923 using linear discriminant analysis (LDA), logistic regression (LR), random forest (RF) and support vector machine (SVM) predictive model analyses. Our work reveals promise for using reverse phase protein arrays for biomarker discovery and validation in serum samples.

  1. RayBiotech Inc, Guangzhou, China
  2. RayBiotech Inc, Parkway Lane, Norcross, GA, USA
  3. Guangdong Provincial Academy of Chinese Medical Sciences, Guangzhou, China
  4. Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, China
  5. The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China
  6. South China Biochip Research Center, Guangzhou, China

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