Antibodies as targeting moieties: affinity measurements, conjugation chemistry and applications in immunoliposomes

Herron, James N.; Gentry, Christine; Davies, Sean S.; Ai-ping, Wei; Jun, Lin

doi:10.1016/0168-3659(94)90162-7

Cited by 12 publications

(7 citation statements)

References 38 publications

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“…The hybridoma cell line for 4-4-20 (Kranz & Voss, 1981a) was obtained from Prof. E. W. Voss, Jr. (University of Illinois at Urbana-Champaign). The 4-4-20 antibody was produced in BALB/c mice and purified as described by Herron et al (1994). The purified antibody was digested with pepsin to produce F(ab')2 fragments as described by Parham (1983).…”

Section: Methodsmentioning

confidence: 99%

Molecular Dynamics of the Anti-Fluorescein 4-4-20 Antigen-Binding Fragment. 2. Time-Resolved Fluorescence Spectroscopy

Lim

Jameson²,

Gentry³

et al. 1995

Biochemistry

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Time-resolved fluorescence experiments were performed to investigate the dynamic aspects of the antigen-binding fragment (Fab) of a high-affinity monoclonal antibody (4-4-20) which binds the fluorescent hapten fluorescein. Both the unliganded Fab and a complex of the Fab with a nonfluorescent analog of fluorescein (fluoresceinamine, FLM) were examined. A fluorescence polarization probe [5-[[2-[(iodoacetyl)amino]ethyl]amino]naphthalene-1-sulfonic acid, AEDANS] was covalently attached to the C-terminus of the Fab. Experiments were performed at three different temperatures (10, 25, and 35 degrees C), and phase-modulation data sets were collected for five different molar ratios of FLM to Fab at each temperature. Global analyses were then used to extract values for fluorescence lifetime and rotational correlation time from these data. In the lifetime analysis the best fit was obtained when the emission of AEDANS was described by a Lorentzian distribution of lifetimes (tau = 15.6 ns, distribution width = 3.4 ns, both at 25 degrees C), which suggested that the probe experienced a heterogeneous environment. Anisotropy analyses suggested that two different rotational components were present. The first was attributed to the global motion of the Fab and exhibited a rotational correlation time (theta 1) of ca. 33 ns at 25 degrees C. This component was relatively unaffected by antigen binding. The second rotational component was attributed to the local or segmental motion within the Fab and exhibited a rotational correlation time (theta 2) of 1.1 ns at 25 degrees C. This value increased by more than 50% upon antigen binding, a result which was consistent with molecular dynamics simulations of the same Fab--fluorescein system [Lim & Herron (1995) Biochemistry 34, 6962-6974]. Furthermore, statistical analysis showed that this increase was significant at the 95% confidence level.

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Section: Methodsmentioning

confidence: 99%

Molecular Dynamics of the Anti-Fluorescein 4-4-20 Antigen-Binding Fragment. 2. Time-Resolved Fluorescence Spectroscopy

Lim

Jameson²,

Gentry³

et al. 1995

Biochemistry

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“…104 Drug carriers can bear either maleimide (Fig. 4a) 41,61,62,[82][83][84][85][86][87][88][89][90][91][92][93][94][95][106][107][108][109] or activated thiol groups (Fig. 4b and c) 60,104,[110][111][112][113] on the surface for functionalization.…”

Section: Maleimide and Thiol Reactionmentioning

confidence: 99%

“…4b and c) 60,104,[110][111][112][113] on the surface for functionalization. Since maleimide groups are relatively stable under synthesis conditions, maleimide-containing molecules have been used during the synthesis of liposomes, 77,78,[82][83][84]89,100,108 nanoparticles, 62 polymeric micelles, 41 or water-soluble polymer-drug conjugates. 106 For example, N-(4-(p-maleimidophenyl)butyryl)phosphatidylethanolamine (MPB-PE) 82,85,108 and N-[(3-maleimide-1-oxopropyl)aminopropyl polyethyleneglycol-carbamyl] distearoylphosphatidyl-ethanolamine (DSPE-PEG-Mal) 77,78,83,88,100 have been widely employed as maleimide-functionalized anchors in the formulation of maleimide-functionalized liposomes.…”

Section: Maleimide and Thiol Reactionmentioning

confidence: 99%

“…114 As thiol groups are very reactive towards various electrophiles and rapidly deteriorate when in solution, it is difficult to introduce sufficient amounts of thiol residues into biomolecules such as antibodies. The most frequently used modification techniques involve antibody activation N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) [83][84][85]108 and N-succinimidyl-S-acetylthioacetate (SATA) 82 followed by a deprotection procedure using reducing reagents such as dithiothreitol (DTT). Alternatively, the endogenous disulfide groups in the hinge region of the immunoglobulin structure can be reduced to generate half-antibody fragments containing active thiol groups.…”

Section: Maleimide and Thiol Reactionmentioning

confidence: 99%

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Organic nanoscale drug carriers coupled with ligands for targeted drug delivery in cancer

Shi

Shoichet

2009

J. Mater. Chem.

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“…Harris et al50 synthesized PEO thiols using tosylated PEO and the thiolation agent sodium hydrosulfide. Herron et al51 have used N ‐succinimidyl‐3‐(2‐pyridyldithio)propionate (SPDP) to thiolate PEO using an amino‐functionalized PEO. However, neither method allowed one‐step direct thiolation, and the formation of disulfide appeared to be a complication.…”

Section: Introductionmentioning

confidence: 99%

Synthesis and characterization of thiol‐terminated poly(ethylene oxide) for chemisorption to gold surface

Brash

2003

J of Applied Polymer Sci

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Thiol-terminated poly(ethylene oxide) (PEO) was synthesized using two different approaches: esterification of terminal hydroxyl groups with mercaptoacetic acid and amidation using N-hydroxysuccinimidyl PEO (NHS-PEO) and cysteine. The reaction of hydroxyl-terminated PEO with mercaptoacetic acid was carried out in boiling toluene. Different thiolated PEOs, including linear PEOs of varying molecular weights and end-group types, and startype PEOs were synthesized. Nuclear magnetic resonance and infrared spectroscopy were used to characterize the products. The reaction kinetics were also briefly investigated. Gel permeation chromatography was used to investigate the relative amounts of the mono-and disubstituted products in the ␣,-dihydroxy PEOs. NHS-PEO was used both to attach terminal thiol groups via reaction with cysteine and to conjugate other amino acids (and potentially any amino-containing molecule) to PEO. Reactions using NHS-PEO were carried out at room temperature in water. The chemisorption of these thiolated PEOs to gold was expected to yield surfaces resistant to biofouling, in particular to unwanted protein adsorption. Chemisorption of amino acid-, peptide-, or protein-terminated PEOs in addition may yield surfaces having specific biological activity. Work on these aspects will be reported elsewhere.

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Antibodies as targeting moieties: affinity measurements, conjugation chemistry and applications in immunoliposomes

Cited by 12 publications

References 38 publications

Molecular Dynamics of the Anti-Fluorescein 4-4-20 Antigen-Binding Fragment. 2. Time-Resolved Fluorescence Spectroscopy

Molecular Dynamics of the Anti-Fluorescein 4-4-20 Antigen-Binding Fragment. 2. Time-Resolved Fluorescence Spectroscopy

Organic nanoscale drug carriers coupled with ligands for targeted drug delivery in cancer

Synthesis and characterization of thiol‐terminated poly(ethylene oxide) for chemisorption to gold surface

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