Goil S, Barnes C, Thibeault D, Truog W E
Department of Pediatrics, The Children's Mercy Hospital, The University of Missouri-Kansas City School of Medicine 64108, USA.
J Chromatogr B Biomed Sci Appl. 1998 Feb 13;705(2):203-11. doi: 10.1016/s0378-4347(97)00523-9.
Many studies employ bronchoalveolar lavage fluid for assessment of biologically active substances secreted from the lung. However, investigators continue to search for a useful reference standard to correct for the inevitable but variable degree of dilution of this fluid. The glycoprotein, soluble secretory component of IgA, may serve as a valid reference protein. We report a simplified method for the purification of secretory component from colostrum. Soluble secretory component was isolated from human colostrum using serial centrifugation, size-exclusion fractionation and ion-exchange chromatography. Secretory component rich fractions were assayed by enzyme immunoassay. They were also evaluated for total amino acid content and distribution and sequence determination with satisfactory agreement with published results. We then demonstrated that soluble secretory component concentration in tracheal aspirate fluid did not correlate with either albumin or with total protein measured in the same samples. Therefore, we conclude that the secretory component of IgA serves as a useful reference marker because its use may avoid errors resulting from leakage of plasma proteins into epithelial lining fluid. Advantages of this method for establishing a standard for secretory component include ready availability of soluble secretory component, simplicity of the method and relative rapidity of the techniques.
许多研究采用支气管肺泡灌洗液来评估肺分泌的生物活性物质。然而,研究人员仍在继续寻找一种有用的参考标准,以校正这种灌洗液不可避免但程度各异的稀释。糖蛋白,即IgA的可溶性分泌成分,可能是一种有效的参考蛋白。我们报告了一种从初乳中纯化分泌成分的简化方法。使用连续离心、尺寸排阻分级分离和离子交换色谱法从人初乳中分离可溶性分泌成分。通过酶免疫测定法对富含分泌成分的级分进行测定。还对它们的总氨基酸含量、分布及序列测定进行了评估,结果与已发表的结果吻合良好。然后我们证明,气管吸出液中可溶性分泌成分的浓度与同一样本中测得的白蛋白或总蛋白均无相关性。因此,我们得出结论,IgA的分泌成分可作为一种有用的参考标志物,因为使用它可避免因血浆蛋白漏入上皮衬液而导致的误差。该方法用于建立分泌成分标准的优点包括可溶性分泌成分易于获得、方法简便以及技术相对快速。