Biomedical Engineering Reference
In-Depth Information
Table3.
Native and affinity-tail bearing enzymes immobilized on metal chelate supports
Enzyme
Support
Ref.
Lactate dehydrogenase
Cu
2+
-IDA-agarose
[154]
Malate dehydrogenase
Cu
+
-IDA-agarose
[154]
Alkaline phosphatase
Cu
2+
/Zn
2+
/Co
2+
-IDA-agarose
[154]
Cu
2+
-IDA-silica
Ribonuclease A
[114]
Lysozy me
Cu
2+
-IDA-silica
[114]
Glucose oxidase
Cu
2+
-IDA-silica
[114]
Ribonuclease B
Cu
2+
-IDA-silica
[114]
Cu
2+
superose HR10/2
Peroxidase (his)
n
[153]
Glucose oxidase (his)
n
Cu
2+
superose HR10/2
[153]
b
-galactosidase (his)
6
Ni
2+
NTA adsorbent
[150]
b
-galactosidase (Ala-His,Gly-His-Arg-Pro)n
Zn
2+
NTA adsorbents
[149]
Cu
2+
/Zn
2+
IDA-Sepharose
Galactose dehydrogenase (his)
5
[152]
Lactose dehydrogenase (his)
4
Cu
2+
/Zn
2+
IDA-Sepharose
[152]
b
-glucouronidase (his)
4
Cu
2+
/Zn
2+
IDA-Sepharose
[152]
ty was retained on Co
2+
-chelate agarose followed by that with Zn
2+
;Cu
2+
supports
were most inferior in this regard.
Anspach and Hasse [155] reported the immobilization of penicillin G amido-
hydrolase from
E. coli
on immobilised metal chelate supports. A number of the
other proteins and enzymes are also known to bind strongly to metal chelate
supports including ribonuclease A and lysozyme [114].
Immobilized metal chelate supports have also been used in the preparation of
affinity sorbents for enzymes,that do not exhibit high affinity for the immobili-
zed metal ions,by binding to immobilized proteins that strongly bind both to
the carriers and to the enzymes. The “sandwich affinity sorbents”,so called
because the affinity ligands is located between the immobilized metal chelate
and the enzyme,have been used principally in the biospecific interaction
chromatography of enzymes and proteins [114]. Con A has been shown bind
strongly to Cu
2+
iminodiacetic acid functions in view of the presence of six
histidine residues in each subunit [156] and retains affinity towards carbohy-
drates and glycoproteins. Infact Con A appears to be oriented in a manner that
facilitated high accessibility of the carbohydrate binding sites for combining
with carbohydrates and glycoproteins [115]. Con A immobilized on metal
chelate supports has been shown to bind a number of enzymes including per-
oxidase and glucose oxidase [114]. Hale et al. [80] demonstrate that antibodies
bind to Co
2+
-IDA-support via metal binding sites located on C- terminal portion
of the heavy chain. This facilitated the orientation of the antibody with antigen
binding site facing away from the support. It was possible to immobilize anti-
bodies in an exchange inert fashion by oxidising the antibody bound Co
2+
to
Co
3+
and all the investigated subclasses of the antibodies could be immobilized
by this procedure [81]. Considerable potential of enzymes immobilized on
immunoaffinity support exist in industry and analysis [157] and the Co
2+
-
chelate supports may be very useful in preparing supports with favourably
oriented antibodies for the purpose.
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