Anti-mucus polyclonal antibody production, purification and linkage to the surface of albumin microspheres

Angela Macadam, Z.B. Shafi, Claire Marriott, G.P. Martin, S.Elizabeth James

Research output: Contribution to journalArticle

Abstract

This aim of this study was to develop a microparticulate based oral drug delivery system, which could prolong gut transit time by binding via specific interactions to the gut mucus layer. Porcine gastric mucus was semi-purified and used as an antigen to raise a polyclonal antiserum in rabbits. The immunoglobulin fraction of this serum was isolated, purified and tested for homogeneity and cross reactivity. High cross-reactivity was displayed when the antiserum was challenged against types of mucus other than that used as an antigen, but no significant cross-reactivity occurred when challenged against some other common macromolecules. The antibody fraction of this serum was covalently linked to three types of albumin microspheres (MS) using 1-ethyl-3(3-dimethyl aminopropyl) carbodiimide. The MS employed had either a hydrophobic, a hydrophilic or a carboxymethylated surface, and were prepared and characterised as described earlier (MacAdam, A.B., Shafi, Z.B., Martin, G.P. and James, S.L. 1997. Preparation of hydrophobic and hydrophilic MS and determination of surface carboxylic acid and amino residues. Int. J. Pharm. 151, 47–55). Binding of these MS to both radioiodinated mucin in suspension and to isolated gut segments was measured. Hydrophilic and carboxymethylated MS with surface-associated antibody bound significantly more mucin from a suspension than did uncoated controls. Similarly, anti-mucus antibody-coated hydrophilic and carboxymethylated MS bound more strongly to an isolated gut segment than did uncoated controls or controls coated in an antibody specific for albumin. These results suggest anti-mucus antibody coated albumin MS may be a useful model to act as comparators in studies aimed at developing drug delivery systems with delayed gastrointestinal transit
Original languageEnglish
Pages (from-to)147-158
Number of pages12
JournalInternational Journal of Pharmaceutics
Volume195
Issue number1-2
DOIs
Publication statusPublished - 15 Feb 2000

Keywords

  • polyclonal antibodies
  • bovine serum albumin
  • albumin microspheres
  • ligand binding
  • antibody production
  • antibody isolation
  • S-35 anti-rabbit IgG
  • antibody linkage
  • ELISA
  • PAGE
  • carboxymethylated MS
  • bioadhesion
  • mucoadhesive strength
  • radio-iodinated mucus
  • perfusion of rat jejunum segments
  • carboxymethylated mucus

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