Chapter 42 : Western Blot Analysis for the Detection of Anti-Glomerular Basement Membrane Antibodies and Anti-Phospholipase A2 Receptor Antibodies

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Western Blot Analysis for the Detection of Anti-Glomerular Basement Membrane Antibodies and Anti-Phospholipase A2 Receptor Antibodies, Page 1 of 2

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Anti-basement membrane antibodies and diffuse alveolar hemorrhage (Goodpasture's syndrome) receives its eponym from Ernest Goodpasture. While studying the influenza pandemic after World War I, Goodpasture studied an 18-year-old male, who died of pulmonary hemorrhage, and diagnosed his pathological findings as systemic vasculitis (1). Although the disease that Goodpasture described is probably different from the disease that now carries his eponym (2), Goodpasture's disease is now well recognized as one of the causes of pulmonary hemorrhage and/or acute renal failure. Immunofluorescence later identified that some cases of pulmonary hemorrhage and renal failure were associated with the linear deposition of immunoglobulins deposited along the pulmonary and glomerular basement membranes (GBM) (3–5). The term Goodpasture's is now generally reserved for the diseases of pulmonary hemorrhage and renal failure with anti-basement membrane antibodies (6, 7).

Citation: Collins A, Smith R. 2016. Western Blot Analysis for the Detection of Anti-Glomerular Basement Membrane Antibodies and Anti-Phospholipase A2 Receptor Antibodies, p 385-390. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch42
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Image of FIGURE 1

Lane 1 shows a Western blot of known positive anti-GBM (1:20) identifying the required bands of 28 and 48 to 50 kDa. Lane 2 shows a negative serum sample. Note the higher-molecular-mass bands in both the positive and negative sera.

Citation: Collins A, Smith R. 2016. Western Blot Analysis for the Detection of Anti-Glomerular Basement Membrane Antibodies and Anti-Phospholipase A2 Receptor Antibodies, p 385-390. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch42
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Image of FIGURE 2

Example of a Western blot for anti-phospholipase A2 receptor antibodies. Lanes 1 and 3 show positive serum samples at 1:20 (lane 1) and 1:100 (lane 3) dilutions from a patient with primary/idiopathic membranous nephropathy (MGN) of the anti-phospholipase A2 receptor type. Lane 2 shows a negative control at 1:20 dilution from an ANCA-positive patient. On the right are molecular mass markers.

Citation: Collins A, Smith R. 2016. Western Blot Analysis for the Detection of Anti-Glomerular Basement Membrane Antibodies and Anti-Phospholipase A2 Receptor Antibodies, p 385-390. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch42
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