Polyclonal‐antibody‐mediated insolubilization and stabilization of papain

Abstract

Antisera raised in rabbits against native iodoacetamide- and iodoacetic acid-modified papain contained precipitating and non-inhibitory antibodies. Papain could be insolubilized as enzyme-antibody adducts with the gamma-globulin fraction derived from the antiserum. Two insolubilized papain preparations, designated A and B, with low and high antibody/enzyme ratios, respectively, exhibited high enzyme activity and markedly enhanced thermal and denaturant stabilities. However, papain antibody adduct B was superior in activity and stability over preparation A. The usefulness of immobilized papain in biopharmaceutical and other industries is also discussed.

Amebic cysteine proteinase 2 (EhCP2) plays either a minor or no role in tissue damage in acute experimental amebic liver abscess in hamsters

“…The stabilizing or enhancing effects of the IgG anti-CP antibodies on the proteolytic activity of both live axenic trophozoites of E. histolytica and affinity-purified CP was an interesting result. Although we have no explanation for it, this phenomenon is observed with other anti-enzyme antibodies (Jafri et al 1993;Khan and Iqbal 2000) and is usually attributed to changes in the enzyme molecule caused by the antibody, which is not combining with the active site of the enzyme. Several amebic molecules have been used as immunogens, in the hope of stimulating the production of antibodies protecting against experimental amebiasis (Cheng et al 1999;Marinets et al 1997;Zhang et al 1994).…”

Stabilization of Bacillus licheniformis α-amylase by specific antibody which recognizes the N-terminal fragment of the enzyme

“…Therefore, most of BLA activity was remained even in complex form, this could be proved by high η values as an evident (0.87); η is the effectiveness factor, and is calculated by the ratio of enzyme activity in the presence and absence of IgG. High retention of activity upon insolubilization of papain [18], stem bromelain [12], carboxypeptidase [37] and leucine aminopeptidase [38] with their antibodies has also been reported, whereas the other enzymes, like aspartate aminopeptidase [39] and plasminogen activator [40], did not show much remained activity under similar conditions.…”

“…Previous attempts were focused on stabilization of enzymes through formation of immuno complex and binding on antibody supports [18][19][20]. At first, we produced a polyclonal antibody against the intact BLA which could completely inhibit the enzyme.…”

“…Both polyclonal [18,19] and monoclonal antibodies [20] have been successfully employed for stabilization and immobilization of enzymes on solid supports [16], or as enzyme-antibody aggregates [21][22][23]. The stability of various enzymes including glucose oxidase, invertase and catalase against inactivation has been remarkably improved using these strategies [18]. Here, a polyclonal antibody against the TF18K was prepared and applied to improve the thermostability of BLA.…”

“…In addition, among large number of strategies that are being currently used to investigate the improvement of [12][13][14][15], the use of specific antienzyme antibodies appears versatile and promising [16,17]. Both polyclonal [18,19] and monoclonal antibodies [20] have been successfully employed for stabilization and immobilization of enzymes on solid supports [16], or as enzyme-antibody aggregates [21][22][23]. The stability of various enzymes including glucose oxidase, invertase and catalase against inactivation has been remarkably improved using these strategies [18].…”