Polyreactivity as an acquired artefact, rather than a physiologic property, of antibodies: evidence that monoreactive antibodies may gain the ability to bind to multiple antigens after exposure to low pH

TY - JOUR

T1 - Polyreactivity as an acquired artefact, rather than a physiologic property, of antibodies

T2 - evidence that monoreactive antibodies may gain the ability to bind to multiple antigens after exposure to low pH

AU - McMahon, Michael J.

AU - O'Kennedy, Richard

PY - 2000/7/31

Y1 - 2000/7/31

N2 - Evidence is presented that monoreactive antibodies exposed to low pH may acquire the ability to bind to multiple antigens. M11, a murine, monoclonal, IgM(K) anti-goat IgG (GIgG) was purified from a hybridoma supernatant by elution at low pH from an anti-mu-Sepharose 4B affinity column. By measuring the specific antiGIgG activities and the affinity constants for the interactions of M11, pre- and post-affinity-purification, with GIgG, M11 was shown to be monoreactive before purification. Quite unexpectedly, however, the affinity-purified M11 reacted extensively with size-fractionated liver proteins when tested in an immunoblot, clearly indicating that it was polyreactive. It was concluded that the exposure to low pH had altered the M11 binding-site so as to allow it to bind to many different proteins. This phenomena provides an alternative basis for interpreting the polyreactivity observed following affinity-purification.

AB - Evidence is presented that monoreactive antibodies exposed to low pH may acquire the ability to bind to multiple antigens. M11, a murine, monoclonal, IgM(K) anti-goat IgG (GIgG) was purified from a hybridoma supernatant by elution at low pH from an anti-mu-Sepharose 4B affinity column. By measuring the specific antiGIgG activities and the affinity constants for the interactions of M11, pre- and post-affinity-purification, with GIgG, M11 was shown to be monoreactive before purification. Quite unexpectedly, however, the affinity-purified M11 reacted extensively with size-fractionated liver proteins when tested in an immunoblot, clearly indicating that it was polyreactive. It was concluded that the exposure to low pH had altered the M11 binding-site so as to allow it to bind to many different proteins. This phenomena provides an alternative basis for interpreting the polyreactivity observed following affinity-purification.

KW - Polyreactive

KW - Immunoblot

KW - Affinity-purification

U2 - 10.1016/S0022-1759(00)00196-4

DO - 10.1016/S0022-1759(00)00196-4

M3 - Article

C2 - 10915844

SN - 0022-1759

VL - 241

SP - 1

EP - 10

JO - Journal of immunological methods

JF - Journal of immunological methods

IS - 1-2

ER -