Abstract
Activation of the contact system has been documented in severe sepsis and hereditary angioedema, but a sensitive, specific, and quantitative assay for assessing the degree of involvement of this proteolytic enzyme cascade is not yet available. We have developed a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) for the α2-macroglobulin-kallikrein (α2M-Kal) complex using an F(ab′)2 derivative of a monospecific polyclonal antibody against α2M as the capture antibody and a unique murine monoclonal antibody, 13G11, against the heavy chain of kallikrein as the detector antibody. The assay does not detect complexes in normal plasma but reacts with complexes generated by activating normal plasma with dextran sulfate at 4°C in a range of 5 to 375 nmol/L. A close correlation of the ELISA with an amidolytic assay for α2M-Kal was documented. Patients with sepsis syndrome but negative bacterial blood cultures did not show elevated plasma complexes, whereas a majority of those with positive blood cultures did show modest elevation and a single patient with septic shock showed a very high level of α2M-Kal complex. Similarly, a patient with classic hereditary angioedema (HAE) showed increased concentration of complexes on three separate occasions during attacks but normal levels between attacks. Two other HAE patients did not show elevated levels at quiescent periods. The ELISA for α2M-Kal appears to be sensitive, specific, and quantitative, and it can be used to reflect the degree of contact system activation in human sepsis and in HAE attacks.
| Original language | English |
|---|---|
| Pages (from-to) | 2660-2667 |
| Number of pages | 8 |
| Journal | Blood |
| Volume | 77 |
| Issue number | 12 |
| State | Published - Jun 15 1991 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Hematology
Cite this
α2-macroglobulin-kallikrein complexes detect contact system activation in hereditary angioedema and human sepsis. / Kaufman, Nathan; Page, Jimmy D.; Pixley, Robin A.; Schein, Roland; Schmaier, Alvin H.; Colman, Robert W.
In: Blood, Vol. 77, No. 12, 15.06.1991, p. 2660-2667.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - α2-macroglobulin-kallikrein complexes detect contact system activation in hereditary angioedema and human sepsis
AU - Kaufman, Nathan
AU - Page, Jimmy D.
AU - Pixley, Robin A.
AU - Schein, Roland
AU - Schmaier, Alvin H.
AU - Colman, Robert W.
PY - 1991/6/15
Y1 - 1991/6/15
N2 - Activation of the contact system has been documented in severe sepsis and hereditary angioedema, but a sensitive, specific, and quantitative assay for assessing the degree of involvement of this proteolytic enzyme cascade is not yet available. We have developed a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) for the α2-macroglobulin-kallikrein (α2M-Kal) complex using an F(ab′)2 derivative of a monospecific polyclonal antibody against α2M as the capture antibody and a unique murine monoclonal antibody, 13G11, against the heavy chain of kallikrein as the detector antibody. The assay does not detect complexes in normal plasma but reacts with complexes generated by activating normal plasma with dextran sulfate at 4°C in a range of 5 to 375 nmol/L. A close correlation of the ELISA with an amidolytic assay for α2M-Kal was documented. Patients with sepsis syndrome but negative bacterial blood cultures did not show elevated plasma complexes, whereas a majority of those with positive blood cultures did show modest elevation and a single patient with septic shock showed a very high level of α2M-Kal complex. Similarly, a patient with classic hereditary angioedema (HAE) showed increased concentration of complexes on three separate occasions during attacks but normal levels between attacks. Two other HAE patients did not show elevated levels at quiescent periods. The ELISA for α2M-Kal appears to be sensitive, specific, and quantitative, and it can be used to reflect the degree of contact system activation in human sepsis and in HAE attacks.
AB - Activation of the contact system has been documented in severe sepsis and hereditary angioedema, but a sensitive, specific, and quantitative assay for assessing the degree of involvement of this proteolytic enzyme cascade is not yet available. We have developed a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) for the α2-macroglobulin-kallikrein (α2M-Kal) complex using an F(ab′)2 derivative of a monospecific polyclonal antibody against α2M as the capture antibody and a unique murine monoclonal antibody, 13G11, against the heavy chain of kallikrein as the detector antibody. The assay does not detect complexes in normal plasma but reacts with complexes generated by activating normal plasma with dextran sulfate at 4°C in a range of 5 to 375 nmol/L. A close correlation of the ELISA with an amidolytic assay for α2M-Kal was documented. Patients with sepsis syndrome but negative bacterial blood cultures did not show elevated plasma complexes, whereas a majority of those with positive blood cultures did show modest elevation and a single patient with septic shock showed a very high level of α2M-Kal complex. Similarly, a patient with classic hereditary angioedema (HAE) showed increased concentration of complexes on three separate occasions during attacks but normal levels between attacks. Two other HAE patients did not show elevated levels at quiescent periods. The ELISA for α2M-Kal appears to be sensitive, specific, and quantitative, and it can be used to reflect the degree of contact system activation in human sepsis and in HAE attacks.
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M3 - Article
C2 - 1710516
AN - SCOPUS:0025848380
VL - 77
SP - 2660
EP - 2667
JO - Blood
JF - Blood
SN - 0006-4971
IS - 12
ER -