Vandermeeren M, Daneels G, Bergers M, van Vlijmen-Willems I, Pol A, Geysen J, Schalkwijk J
Janssen Research Foundation, Janssen Pharmaceutica, Beerse, Belgium.
Arch Dermatol Res. 2001 Jul;293(7):343-9. doi: 10.1007/s004030100238.
SKALP/elafin is an epithelial proteinase inhibitor with antimicrobial properties that is not normally expressed in human epidermis, but is induced under inflammatory conditions and in some types of skin cancer. SKALP is a member of the recently described trappin gene family, which encodes a new class of proteins, characterized by a four-disulphide core and a transglutaminase substrate domain. Polyclonal antisera against SKALP have been shown to be useful for monitoring disease activity in psoriasis and tumour differentiation in squamous cell carcinoma. We developed ten different mouse monoclonal antibodies (mAbs) against synthetic peptides corresponding to a hexapeptide epitope in the transglutaminase substrate domain and three mAbs recognizing an epitope in the proteinase-inhibiting domain. The antibodies could be used with high specificity by immunohistochemistry on formalin-fixed tissue, by affinity chromatography, by Western blotting, and by enzyme-linked immunoadsorbent assay (ELISA) for the detection of SKALP/elafin. These antibodies have several advantages over existing polyclonal antisera, such as a defined epitope, the detection of full-length SKALP/elafin and unlimited supply. An antibody against the hexapeptide epitope, which is common to all known human, simian, bovine and swine trappin family members, was used to immunolocalize bovine trappins expressed in trachea, that have recently been discovered. These mAbs will serve as important new tools to measure SKALP/elafin and trappin family members in research and diagnostics.
SKALP/弹性蛋白酶抑制因子是一种具有抗菌特性的上皮蛋白酶抑制剂，通常不在人类表皮中表达，但在炎症条件下以及某些类型的皮肤癌中会被诱导表达。SKALP是最近描述的捕集蛋白基因家族的成员，该家族编码一类新的蛋白质，其特征是具有一个四硫键核心和一个转谷氨酰胺酶底物结构域。已证明针对SKALP的多克隆抗血清可用于监测银屑病的疾病活动和鳞状细胞癌的肿瘤分化。我们针对转谷氨酰胺酶底物结构域中的一个六肽表位对应的合成肽制备了十种不同的小鼠单克隆抗体（mAb），以及三种识别蛋白酶抑制结构域中一个表位的单克隆抗体。这些抗体可通过对福尔马林固定组织进行免疫组织化学、亲和层析、蛋白质印迹和酶联免疫吸附测定（ELISA）以高特异性用于检测SKALP/弹性蛋白酶抑制因子。与现有的多克隆抗血清相比，这些抗体具有几个优点，例如明确的表位、全长SKALP/弹性蛋白酶抑制因子的检测以及无限供应。一种针对所有已知人类、猿猴、牛和猪捕集蛋白家族成员共有的六肽表位的抗体，被用于对最近发现的在气管中表达的牛捕集蛋白进行免疫定位。这些单克隆抗体将成为在研究和诊断中测量SKALP/弹性蛋白酶抑制因子和捕集蛋白家族成员的重要新工具。
