title: Highly sensitive detection of cancer antigen human epidermal growth factor receptor 2 using novel chicken egg yolk immunoglobulin.
abstract: Human epidermal growth factor receptor 2 (HER2) is an important biomarker that plays a crucial role in therapeutic decision-making for breast cancer patients. Ensuring the accuracy and reproducibility of HER2 assays by enzyme-linked immunosorbent assay (ELISA), western blot and immunohistochemistry (IHC) requires high sensitive and specific antibodies. Immunoglobulin Y (IgY) is a kind of avian antibody usually isolated from chicken egg yolks. Generation and use of IgY is of increasing interest in a wide variety of applications within the life sciences. In this study, IgY antibodies against two different truncated proteins of the extracellular domain (ECD) of human HER2 were produced, their sensitivity and specificity were evaluated. Specific IgYs were produced by hens immunized with the ECD proteins of human HER2 in long-standing immunization response and were isolated from yolks with a purity of 90% by water dilution, salt precipitations and ultrafiltration. The anti-HER2 IgYs were analytically validated for specificity by ELISA, western blot, immunocytochemistry and IHC. The IgYs bound desired targets in cells and fixed tissues and showed high affinity to HER2. The results demonstrated the viability of detection of HER2 with IgYs and showed promise for the using of IgYs in strict clinical validation.
authors.affiliation: College of Pharmacy, Dalian Medical University, Dalian, Liaoning Province, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Clinical Medicine of Seven-Year-Program, Dalian Medical University, Dalian, Liaoning Province, China.
authors.forename: Yiheng
authors.initials: Y
authors.lastname: Yang
authors.affiliation: The Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning Province, China.
authors.forename: Lifen
authors.initials: L
authors.lastname: Wang
authors.affiliation: The Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning Province, China.
authors.forename: Li
authors.initials: L
authors.lastname: Lv
authors.affiliation: The Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning Province, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhu
authors.affiliation: College of Pharmacy, Dalian Medical University, Dalian, Liaoning Province, China.
authors.forename: Wenqi
authors.initials: W
authors.lastname: Han
authors.affiliation: College of Pharmacy, Dalian Medical University, Dalian, Liaoning Province, China.
authors.forename: Enxia
authors.initials: E
authors.lastname: Wang
authors.affiliation: College of Pharmacy, Dalian Medical University, Dalian, Liaoning Province, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Guo
authors.affiliation: College of Pharmacy, Dalian Medical University, Dalian, Liaoning Province, China. Electronic address: zhenyhwaner@aliyun.com.
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Zhen
mesh_terms: D000818:Animals; D000912:Antibodies, Neoplasm / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D002645:Chickens / Q000276:immunology*; D004527:Egg Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007136:Immunoglobulins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D061986:MCF-7 Cells; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology*
chemical_list: D000912:Antibodies, Neoplasm; D004527:Egg Proteins; D007136:Immunoglobulins; D018719:Erb-b2 Receptor Tyrosine Kinases; C016512:IgY; C508053:ERBB2 protein, human
keywords: Breast cancer; Detection; HER2; IHC; IgY