Sola J P, Pedreño Y, Cerezo A, Peñalver-Mellado M
Probelte Pharma S.L.U., Spain; Doctoral school of the National University of Distance Education (UNED) of Spain, Spain.
Probelte Pharma S.L.U., Spain.
Allergol Immunopathol (Madr). 2018 Sep-Oct;46(5):491-498. doi: 10.1016/j.aller.2017.12.003. Epub 2018 Jan 17.
Allergy to cats is a frequent cause of sensitization to indoor allergens and currently there are few alternatives to specific immunotherapy with cat native extracts. The objective is to develop and characterize a new allergoid to increase the tools available for use in clinical practice.
The allergoid cat dander extract (ACD) was developed from a native cat dander extract (NCD) by modification with glutaraldehyde, and the optimal process control was determined by SDS-PAGE, DOT BLOT and determination of free amine groups. The ACD was characterized in protein profile by SDS-PAGE, size exclusion chromatography (SEC) and peptide footprint. The allergenic profile of ACD was determined by immunoblot, IgE CAP inhibition and IgG competition ELISA. The major allergen content in NCD was obtained by the ELISA sandwich protocol and was extrapolated to ACD.
The control process determined the optimal development of the allergoid. The ACD obtained contains 182.28μg/mg of protein and 11.90μg/mg of Fel d 1. SDS-PAGE and SEC confirmed the presence of high molecular weight proteins in ACD, and the peptide footprint showed the presence of Fel d 1 and Fel d 7. The high degree of polymerization was evidenced with the determination of the reduction of lysine residues in the allergoid, resulting 91.96%. The ACD showed a significant loss of allergenicity respect to NCD, while the IgG-binding capacity was maintained.
The ACD obtained presents a good safety profile, so would be a good alternative for treatment of cat allergy.
对猫过敏是室内过敏原致敏的常见原因，目前除了用猫天然提取物进行特异性免疫疗法外，几乎没有其他选择。目的是开发并表征一种新的变应原，以增加临床实践中可用的工具。
变应原猫皮屑提取物（ACD）由天然猫皮屑提取物（NCD）经戊二醛修饰而成，通过SDS-PAGE、斑点印迹和游离胺基团测定确定最佳工艺控制。通过SDS-PAGE、尺寸排阻色谱（SEC）和肽足迹对ACD进行蛋白质谱表征。通过免疫印迹、IgE CAP抑制和IgG竞争ELISA测定ACD的变应原谱。通过ELISA夹心方案获得NCD中的主要变应原含量，并外推至ACD。
对照过程确定了变应原的最佳开发。获得的ACD含有182.28μg/mg的蛋白质和11.90μg/mg的猫过敏原1（Fel d 1）。SDS-PAGE和SEC证实ACD中存在高分子量蛋白质，肽足迹显示存在Fel d 1和Fel d 7。变应原中赖氨酸残基减少量的测定证明了高度聚合，结果为91.96%。与NCD相比，ACD的变应原性显著降低，而IgG结合能力得以维持。
获得的ACD具有良好的安全性，因此将是治疗猫过敏的良好选择。
