Monoconjugate enzyme linked immunoassay

Pillai, S C; Bachhawat, B. K.

doi:10.1016/0014-5793(78)80295-6

Cited by 12 publications

(7 citation statements)

References 18 publications

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“…We suggested that the large molecular sizes of urease [6] and IgG [7] made it sterically impossible for more than one molecule of these proteins to be coupled to a single molecule of immobilised invertase. Invertase is a glycoprotein containing 50% by wt of carbohydrate and has 60 lysine residues [ 1 I].…”

Section: Resultsmentioning

confidence: 99%

“…One of the primary limitations of enzyme-linked assays even for large macromolecular antigens was the absence of a technique for the reproducible stoichiometric synthesis of enzyme-antigen and enzyme-antibody conjugates. This limitation can largely be overcome by the use of matrix conjugation techniques [6,7]. The proven stability of glutaraldehyde conjugates [ 121 particularly so in comparison with reagents which couple proteins through sulphydryl groups [ 131 makes the use of this bifunctional dialdehyde a preferable alternative.…”

Section: ~Isevierlnorth-hollarld Biomedical Pressmentioning

confidence: 99%

“…The rzsI radioactivity in the precipitates and supernatants of each tube and the invertase activity in the supernatants of (i) and (ii) were checked. We suggested that the large molecular sizes of urease [6] and IgG [7] made it sterically impossible for more than one molecule of these proteins to be coupled to a single molecule of immobilised invertase. Invertase is a glycoprotein containing 50% by wt of carbohydrate and has 60 lysine residues [ 1 I].…”

Section: ~Isevierlnorth-hollarld Biomedical Pressmentioning

confidence: 99%

“…The reversible imrnobilisation of a protein at a critically low density (below which intermolecular conjugation of the immobilised protein cannot occur) on a suitable affinity matrix greatly facilitated the formation of protein-protein monoconjugates between two enzymes [6] and between an enzyme and an antibody [7]. This matrix principle has been extended to the conjugation of a radiolabelled peptide to an enzyme, and the stoichiometry of the resultant conjugate was analysed.…”

Section: Introductionmentioning

confidence: 99%

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Stoichiometric enzyme‐peptide conjugation of an affinity matrix

Pillai¹,

Bachhawat

1979

FEBS Letters

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Section: Resultsmentioning

confidence: 99%

Section: ~Isevierlnorth-hollarld Biomedical Pressmentioning

confidence: 99%

Section: ~Isevierlnorth-hollarld Biomedical Pressmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Stoichiometric enzyme‐peptide conjugation of an affinity matrix

Pillai¹,

Bachhawat

1979

FEBS Letters

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“…The information derivable from these methods with regard to three-dimensional protein structure is, however, limited and often ambiguous. The immobilization of a protein at a critically low density on an affinity matrix (Pillai & Bachhawat, 1977, 1978 …”

Section: (Received 29 September 1980/accepted 14 October 1980)mentioning

confidence: 99%

Chemical cross-linking of a dimeric protein on a modified lectin matrix. A general probe for the chemical topology of oligomeric glycoproteins

1982

Biochemical Journal

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A dimeric glycoprotein, glucose oxidase, was allowed to react with lysine-specific cross-linkers, both when immobilized on a succinoylated lectin matrix at a critically low density and also at a high density in solution. Analysis of the cross-linked complexes thus obtained led to the following inferences with regard to the structure of this protein.(1) Of the 15 lysine residues on each glucose oxidase protomer, none is available on the non-interfacial surfaces. (2) Assuming that this protein possesses C2 symmetry with isologous bonding between subunits, it may be inferred that on each protomer there are at least two lysine clusters along or close to the interprotomeric interface. (3) These 'interfacial' lysine residues on each protomer are so oriented that the E-amino groups of lysine residues a and b on protomer 1 'face', and are very close to, the E-amino groups of lysine residues b' and a' respectively on protomer 2. General inferences on the geometry of dimeric proteins derivable from an analysis of the cross-linked complexes obtained (as well as those not seen) by using this low-density matrix cross-linking approach were enumerated. Modified lectin matrices may prove useful in studying the threedimensional structure of glycoproteins, particularly non-crystallizable oligomers.

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References

Torchilin

1991

Progress in Clinical Biochemistry and Medicine

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Monoconjugate enzyme linked immunoassay

Cited by 12 publications

References 18 publications

Stoichiometric enzyme‐peptide conjugation of an affinity matrix

Stoichiometric enzyme‐peptide conjugation of an affinity matrix

Chemical cross-linking of a dimeric protein on a modified lectin matrix. A general probe for the chemical topology of oligomeric glycoproteins

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