Fab fragments using

Motta-Hennessy, C., Eccles, S.A., Dean, C., and Coghlan, G. (1985) Preparation of 67Ga-labeled human IgG and its Fab fragments using deferoxamine as chelating agent. Eur. J. Nucl. Med. 11, 240-245. 

This chapter describes the procedures developed in the laboratory of Martin Glennie at the Lymphoma Research Unit, Tenovus Laboratory, Southampton, UK, for preparing bispecific F(ab)2 and F(ab)3 and tnspecific antibody (TsAb) F(ab)3 derivatives from antibody Fab fragments using the crosslinker o-phenylenedi-maleimide (o-PDM)... 

We have also developed a chemistry that allows us to attach the Fab to only the inner surfaces of the nanotubes. While still within the pores of the template membrane, the inner surfaces were treated with 3-aminopropyltrimethoxysilane. The template membrane was then dissolved and the amino sites on the inner surfaces were coupled to free amino groups on the Fab fragment using the well-known glutaraldehyde coupling reaction [4]. When 18 mg of these interior-only Fab-modified nanotubes were incubated with 1 mL of a 10 pM racemic mixture of the drug, 80% of the RS (and none of the SR) enantiomer was extracted. 

Fiffire 3. Conjugation of radioiodinated HML to Fab fragments using 2-iminothiolane. 

The mass spectrometry of diazo compounds was reviewed by Zeller (1983) and by Lebedev (1991). It is difficult to record mass spectra of diazonium salts using conventional techniques. With the water thermospray method, however, Schmelzeisen-Redeker et al. (1985) observed the diazonium ion and various fragments such as [Ar+ - N2 + 2H]+ and [Ar + N2 + H20]+. Ambroz et al. (1988) applied the fast atom bombardment (FAB) technique using a 3-nitrobenzylalcohol matrix. Peaks for ArNJ, Ar+, and [M + ArN2]+ and further peaks due to solvated ions were found. 

Fab fragments (M.W. 50,000) have several advantages over IgG (M.W. 150,000) for use as the immunotherapeutic reagent (Smith et al. 1979). The Fab fragments distribute more rapidly and extensively than intact IgG. In addition, they are catabolized and excreted earlier than intact IgG. A final important advantage is that the Fab fragments are less antigenic than the intact IgG. 

C) was also used In the anti-PCP Fab fragment studies discussed below. The inset figure shoes a schematic representation of PCP in the dog as a two-... 

The affinity and cross-reactivity of the whole serum and Fab fragments were determined using equilibrium dialysis for the affinity determination and RIA for the cross-reactivity studies. The average intrinsic affinity constant (Ko) of the antibody (Nisonoff and Pressman 1958) changed very little throughout the... 

SIAB and sulfo-SIAB have been used to make a high-capacity RNA affinity column for the purification of human IRP1 and IRP2 (Allerson et al., 2003), to couple antibodies or Fab fragments to amine-modified microparticles (Harma et al., 2000), and in the attachment of oligonucleotides to surfaces for detection arrays (Adessi et al., 2000). 

Figure 20.13 The thiolation reagent SATA can be used to create sulfhydryl groups on Fab fragments. After deprotection of the acetylated thiol of SATA with hydroxylamine, conjugation with a maleimide-activated enzyme can take place, producing thioether linkages.

Figure 20.14 Periodate oxidation of HRP creates aldehyde groups on the carbohydrate chains of the enzyme. Reaction with a Fab fragment then may be done using reductive amination to produce a lower-molecular-weight complex than would be obtained using intact IgG antibodies.

---