Characterization of morphine-specific monoclonal antibodies showing minimal cross-reactivity with codeine

Usagawa, Takashi; Itoh, Y.; Hifumi, Emi; Takeyasu, Akira; Nakahara, Yoshiaki; Uda, Takaaki

doi:10.1016/0022-1759(93)90080-q

Cited by 25 publications

(15 citation statements)

References 12 publications

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“…The result of SDS-PAGE showed that 41S-2-L was partly cleaved at around 100 h, giving the degraded fragment, 22.5, 21.5, 16.5, and 14.5 kDa bands. On the other hand, the light chain of MA-2 mAb (IgG 1 (n) : antimethamphetamine mAb (Usagawa et al, 1989(Usagawa et al, , 1993Uda et al, 1997)) and human serum albumin were hardly degraded by i41SL1-2-L. i41SL1-2-L exhibited high specificity to protein, which was similar to the result we observed previously (Hifumi et al, 1999(Hifumi et al, , 2000a.…”

Section: Degradation Of Cdrl-1 Peptide By the Light Chain Of I41sl1-2supporting

confidence: 91%

Catalytic features of monoclonal antibody i41SL1‐2 subunits

Hifumi

Kondo

Mitsuda

et al. 2003

Biotech & Bioengineering

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A monoclonal antibody (mAb), i41SL1-2, was obtained by immunizing the peptide of complementarity-determining region-1 (CDRL-1: RSSKSLLYSNGNTYLY) of a super catalytic antibody light chain, 41S-2-L, capable of enzymatically destroying the gp41 molecule of the HIV-1 envelope. From the DNA and the deduced amino acid sequences of the light and heavy chain of i41SL1-2 mAb, molecular modeling was conducted that suggested that both subunits of i41SL1-2 mAb possess catalytic triads in their structures. Especially the light chain of i41SL1-2 mAb possesses a characteristic catalytic triad composed of Asp(1), Ser(27A), and His(93), whose positions are identical to the catalytic antibody light chain, VIPase, of S. Paul and colleagues (see text). The antibody gene of i41SL1-2 light chain and VIPase belong to the same germline, bd2, suggesting that the discrete germline inherently possesses catalytic activity. Both light and heavy chains of i41SL1-2 mAb degraded the antigenic peptide CDRL-1 within 47 and 57 h, respectively. The catalytic reaction constant (kcat) of the light and heavy chain was 6.1 x 10(-1) and 6.2 x 10(-1) min(-1), respectively. These are high values for the natural catalytic antibodies reported so far. The catalytic efficiency (kcat/Km) of the light and heavy chain was 3.1 x 10(5) and 4.9 x 10(4) M(-1) min(-1), respectively. The first cleaved bond of the antigenic peptide by subunits of i41SL1-2 mAb was between Arg(1) and Ser(2) in the sequence of CDRL-1, suggesting a serine protease character.

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Section: Degradation Of Cdrl-1 Peptide By the Light Chain Of I41sl1-2supporting

confidence: 91%

Catalytic features of monoclonal antibody i41SL1‐2 subunits

Hifumi

Kondo

Mitsuda

et al. 2003

Biotech & Bioengineering

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“…(28)(29)(30) MAbs against morphine are also reported; these are of various specificities. (18)(19)(20) In these type of strategies, sometimes chemical modifications of hapten molecule, as well as modification of antibody-producing cells, are taken into consideration together. Because of these types of modifications, Usagawa et al (28) could obtain antibodies of high specificities even for codeine, as reported to exhibit negligible cross-reaction.…”

Section: Discussionmentioning

confidence: 99%

“…(15) Reports on polyclonal and monoclonal antibodies (MAbs) against morphine are available. (16)(17)(18)(19)(20) In these, specificity and affinity are of prime importance. In this study, we have produced a specific and high-affinity antibody against morphine and characterized them with a novel ELISA using penicillinase as label that was previously reported for a number of hapten molecules.…”

Section: Introductionmentioning

confidence: 99%

Preparation and Characterization of Specific and High-Affinity Monoclonal Antibodies Against Morphine

Rahbarizadeh

Rasaee

Madani³

et al. 2000

Hybridoma

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A C6-hemisuccinate derivative of morphine was prepared and conjugated to bovine serum albumin. High titer antibody producing spleen cells were removed and fused with myeloma cells of Sp2/0 origin. A C3-hemisuccinate derivative of morphine was prepared and conjugated to enzyme penicillinase used as a tracer molecule. A novel enzyme-linked immunoadsorbent assay was developed using this conjugate to screen and characterize the monoclonal antibody produced in these experiments. After two successive limiting dilutions, antibodies produced by 5 clones with good affinities ranging from 10(8) to 10(12) M(-1) and less cross-reaction (least for codeine and other structurally related molecules) were selected. These clones were found to be of IgG class with kappa light chain. Subclass determination showed that two of the clones produced IgG2b and three of them produced IgG1 type of antibody. Affinity purifications were performed for the selected clone (MOR-I). Purified antibody was coated onto the wells of microtiter plate. The standard curve was constructed with a sensitivity of 100 pg/mL covering up to 10 ng/mL in buffer and urine. The slope of the standard curve for selected clone in buffer and urine was calculated to be -0.7 and -0.64, respectively.

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“…For example, Glasel and Sawada reported that their antibody cross-reacted with codeine [4,5]. Usagawa et al reported that their antibody exhibited cross-reactivity with nalorphine, morphine-6glucuronide and naltrexone, but not with codeine [9]. Rahbarizadeh et al [12] established an antibody that had 10% cross-reactivity with heroin and low cross-reactivity with codeine.…”

Section: Introductionmentioning

confidence: 93%

“…After a polyclonal anti-morphine antibody was reported in 1971, a number of studies attempted to establish anti-morphine antibodies with higher specificities [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22]. However, all of these antibodies have shown cross-reactivity among structurally related compounds, particularly with codeine and heroin.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a Single Chain Fv Antibody that Reacts with Free Morphine

Matsukizono

Kamegawa²,

Tanaka

et al. 2013

Antibodies

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An immune phage library derived from mice, hyperimmunized with morphine-conjugated BSA, was used to isolate a single-chain Fv (scFv) clone, M86, with binding activity to morphine-conjugated thyroglobulin (morphine-C-Tg) but not to codeine-, cocaine-, or ketamine-conjugated Tg. Surface plasmon resonance analysis using a morphine-C-Tg-coupled CM5 sensor chip showed that the Kd value was 1.26 × 10−8 M. To analyze its binding activity to free morphine and related compounds, we performed a competitive ELISA with M86 and morphine-C-Tg in the absence or presence of varying doses of free morphine and related compounds. IC50 values for opium, morphine, codeine, and heroin were 257 ng/mL, 36.4, 7.3, and 7.4 nM, respectively. Ketamine and cocaine exhibited no competitive binding activity to M86. Thus, we established a phage library-derived scFv, M86, which recognized not only free morphine and codeine as opium components but also heroin. This characteristic of M86 may be useful for developing therapeutic reagents for opiate addiction and as a free morphine-specific antibody probe

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Characterization of morphine-specific monoclonal antibodies showing minimal cross-reactivity with codeine

Cited by 25 publications

References 12 publications

Catalytic features of monoclonal antibody i41SL1‐2 subunits

Catalytic features of monoclonal antibody i41SL1‐2 subunits

Preparation and Characterization of Specific and High-Affinity Monoclonal Antibodies Against Morphine

Characterization of a Single Chain Fv Antibody that Reacts with Free Morphine

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