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Detection and Quantification of MUC7 in Submandibular, Sublingual, Palatine, and Labial Saliva by Anti-peptide Antiserum
J.G.M. Bolscher
Academic Centre for Dentistry Amsterdam, Department of Oral Biochemistry, Van der Boechorststraat 7, NL-1081 BT Amsterdam, The Netherlands
J. Groenink
Academic Centre for Dentistry Amsterdam, Department of Oral Biochemistry, Van der Boechorststraat 7, NL-1081 BT Amsterdam, The Netherlands
J.S. van der Kwaak
Academic Centre for Dentistry Amsterdam, Department of Oral Biochemistry, Van der Boechorststraat 7, NL-1081 BT Amsterdam, The Netherlands
P.A.M. van den Keijbus
Academic Centre for Dentistry Amsterdam, Department of Oral Biochemistry, Van der Boechorststraat 7, NL-1081 BT Amsterdam, The Netherlands
W. van't Hof
Academic Centre for Dentistry Amsterdam, Department of Oral Biochemistry, Van der Boechorststraat 7, NL-1081 BT Amsterdam, The Netherlands
E.C.I. Veerman
Academic Centre for Dentistry Amsterdam, Department of Oral Biochemistry, Van der Boechorststraat 7, NL-1081 BT Amsterdam, The Netherlands
A.V. Nieuw Amerongen
Academic Centre for Dentistry Amsterdam, Department of Oral Biochemistry, Van der Boechorststraat 7, NL-1081 BT Amsterdam, The Netherlands
The large carbohydrate moiety of low-Mr salivary mucin MUC7 (originally referred to as MG2) is subject to variations. Biochemical analysis and quantification of MUC7 in saliva samples require recognition tools that are independent of the carbohydrate moiety. Therefore, we have evoked three antisera to synthetic peptides of MUC7. One of these (CpMG2), raised against the C-terminal peptide, recognized native MUC7 in saliva and was characterized further. Recognition of MUC7 by CpMG2 turned out to be specific, resistant to dissociating and reductive treatments, and independent of glycosylation differences, as indicated by Western analysis and ELISA. The antiserum could be used to monitor MUC7 during purification procedures. MUC7 was demonstrated in small volumes of saliva from all (sero)mucous glands, including the palate and lip. Analysis with antibodies and lectins indicated large variations in amount as well as in glycosylation of MUC7. An ELISA was developed to determine the relative quantity of MUC7 in the glandular salivas: mean values of approximately 220, 980, and 100 µg mucin per mL were found in submandibular, sublingual, and palatine saliva, respectively.
Key Words: antisera • mucin • MG2 • MUC7 • synthetic peptide
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Journal of Dental Research, Vol. 78, No. 7,
1362-1369 (1999)
DOI: 10.1177/00220345990780071101
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