Simplified method for purification of colostrum to obtain secretory component of immunoglobulin A, using secretory component as a reference protein in tracheal aspirate fluid.
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.
Goil
S
S
Department of Pediatrics, The Children's Mercy Hospital, The University of Missouri-Kansas City School of Medicine 64108, USA.
Barnes
C
C
Thibeault
D
D
Truog
W E
WE
D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange; D003126:Colostrum / Q000737:chemistry*; D006801:Humans; D007124:Immunoenzyme Techniques; D007070:Immunoglobulin A / Q000737:chemistry*; D007231:Infant, Newborn; D012015:Reference Standards; D012635:Secretory Component / Q000302:isolation & purification*; D013396:Suction; D014132:Trachea / Q000378:metabolism*
D007070:Immunoglobulin A; D012635:Secretory Component