Bioaffinity Based Immobilization of Enzymes - Advances in Biochemical Engineering Biotechnology - page 202

bilization of various enzymes that can be linked either to the antigen or the anti-
body [13]. Considerable know-how regarding the immunization,isolation of
specific antibodies and their immobilization has emerged principally in connec-
tion with the immunoaffinity purification of enzymes and this can be readily
adopted for immunoaffinity immobilization of various enzymes.

2.1
Monoclonal or Polyclonal Antibodies?

Both monoclonal and polyclonal antibodies have been employed in the immo-
bilization of enzymes and their relative merits and limitations examined
(Table 2). Hybridoma cultures can provide a spectrum of monoclonal anti-
bodies from which appropriate population can be conveniently screened and
their continuous supply ensured from the selected hybridoma clones [17].
Culturing of hybridomas continues to however remain an expensive endeavour
due to the high costs of culture media and experimental set up required. Poly-
clonal antibodies on the other hand can be relatively inexpensive especially if
they can be raised in a large animal like pig or goat. Heterogeneity of the poly-

Table2. Enzymes immobilized favourably with the help of antibodies

Enzyme Antibody Support matrix Stabilization against Ref.

b -Galactosidase PC - Temperature [14]

Gulonolactone oxidase PC - In vivo Proteolysis [15]

Transglutaminase PC/MC Agarose beads Temperature [16]

Carboxy peptidase A MC Eupergit C Temperature,17, 19]

pH,storage in cold

Lactate dehydrogenase MC Eupergit C Storage [19]

Glucose oxidase MC Sepharose Recycling [13,20]

Chymotrypsin PC Sepharose - [21]

Subtilisin PC - Sodium hypochloride [22]

Temperature

Nitrate reductase MC Sepharose Temperature [23]

Alpha amylase PC - Temperature,lyophlization [22, 24]

Freezing & Thawing

Glucoamylase PC - Temperature,Ethanol [22,24]

Horse radish peroxidase MC - - [25]

Trypsin PC Sepharose - [26]

Urease PC Nylon pH [27]

NAD glycohydrolase PC Nylon Storage [27]

Invertase PC Sepharose Temperature [28]

Glycosyl
Specific
PC Sepharose Temperature [29,30]

L -Hydantoinase PC Sepharose Incubation [31]

PC -Polyclonal; MC -Monoclonal.