Eydoux Cécilia, Aloulou Ahmed, De Caro Josiane, Grandval Philippe, Laugier René, Carrière Frédéric, De Caro Alain
Laboratoire d'Enzymologie Interfaciale et de Physiologie de la Lipolyse, UPR 9025, CNRS-Institut de Biologie Structurale et Microbiologie, 31, Chemin Joseph-Aiguier, 13402 Marseille Cedex 20, France.
Biochim Biophys Acta. 2006 Oct;1760(10):1497-504. doi: 10.1016/j.bbagen.2006.06.005. Epub 2006 Jun 17.
Human pancreatic lipase-related protein 2 (HPLRP2) was previously found to be secreted by the exocrine pancreas. HPLRP2 shows a high level of activity on galactolipids, and might be involved in the digestion of these common vegetable lipids. Specific antibodies were raised in rabbits using a synthetic HPLRP2 peptide selected for its weak amino acid homology with the corresponding peptides of classical human pancreatic lipase (HPL) and human pancreatic lipase-related protein 1 (HPLRP1). ELISA and Western blotting data showed that these antibodies did not react with HPL or HPLRP1. Various tissues from the digestive tract were subjected to Western blotting analysis with the specific anti-peptide HPLRP2 antibody and the expression of HPLRP2 was detected in the pancreas and colon. An ELISA was developed for specifically measuring the HPLRP2 levels in pure pancreatic juice. This procedure was performed using the anti-peptide HPLRP2 antibody as the captor antibody and a biotinylated anti-HPLRP2 polyclonal antibody as the detector antibody. The lowest HPLRP2 quantification limit was found to be 50 microg/L and the reference range for the present assay was 50 microg-500 microg/L. HPL and HPLRP2 levels were measured using specific ELISAs in pancreatic juice from patients with and without pancreatic disorders. Patients with chronic calcifying pancreatitis (CCP) had significantly lower levels of both HPL and HPLRP2 than the controls subjects. The mean HPLRP2 to HPL ratio was estimated to be 28.30% (w/w) and 23.96% (w/w) in controls subjects and CCP patients, respectively, and the difference was not significant. The levels of HPL and HPLRP2 are therefore similarly reduced in both healthy patients and CCP patients.
人胰腺脂肪酶相关蛋白2（HPLRP2）先前被发现由胰腺外分泌部分泌。HPLRP2对半乳糖脂具有高水平活性，可能参与这些常见植物脂质的消化。利用一种合成的HPLRP2肽在兔体内制备特异性抗体，该肽因其与经典人胰腺脂肪酶（HPL）和人胰腺脂肪酶相关蛋白1（HPLRP1）的相应肽段氨基酸同源性较弱而被选用。酶联免疫吸附测定（ELISA）和蛋白质印迹数据表明，这些抗体不与HPL或HPLRP1发生反应。用特异性抗肽HPLRP2抗体对来自消化道的各种组织进行蛋白质印迹分析，在胰腺和结肠中检测到HPLRP2的表达。开发了一种ELISA方法专门用于测定纯胰液中HPLRP2的水平。该检测过程使用抗肽HPLRP2抗体作为捕获抗体，生物素化的抗HPLRP2多克隆抗体作为检测抗体。发现HPLRP2的最低定量限为50微克/升，本检测方法的参考范围为50微克至500微克/升。使用特异性ELISA测定有或无胰腺疾病患者胰液中的HPL和HPLRP2水平。慢性钙化性胰腺炎（CCP）患者的HPL和HPLRP2水平均显著低于对照组。对照组和CCP患者中HPLRP2与HPL的平均比值分别估计为28.30%（重量/重量）和23.96%（重量/重量），差异不显著。因此，健康患者和CCP患者中HPL和HPLRP2的水平均同样降低。
