. 2020 Dec 13;12(12):795. doi: 10.3390/toxins12120795

Table 2.

Home-made ISs for the analysis of toxins.

Target Toxin(s)	Matrix	Extraction Solvent; Dilution Factor and Solvent	Sorbent, Amount of Abs	Grafting Yield or Density; Capacity	Extraction Mode	V_sample (eq. Sample Amount)/ Amount of Sorbent	Washing	Elution	Analysis	Ref.
Toxins with MW < 1500
Bacterial toxin
TTX	Marine organisms	MeOH, 1% AA; PBS (20% MeOH)	CNBr-Sepharose (0.5 g); mAbs (6 mg)	1106 ng/mL; -	Off-line SPE	25 mL (eq. 1 g)/0.5 g	Water	MeOH, AA (1%)	LC/MS–MS	[69]
Phycotoxins–Cyanotoxins
Anatoxin-a	Pure water	-	NHS-Sepharose beads (27 µm, 10 µL); mAbs (100 µg)	-; 20 ng	dSPE	20 mL/ 10 µL	-	2-propanol	IMS	[70]
MC-LR	Algae extracts	-	Poly(APTES-co-TEOS) monolith; pAbs	-; 0.38 pmol (2.1 µg/g sorbent)	On-line SPE	150 nL/ 45 × 0.1 mm i.d. capillary	PBS	ACN/water (LC mobile phase)	Nano-LC/UV	[71]
	Urine	-	streptavidin-magnetic beads; Biotin-Abs		dSPE	100 µL	-	Water/ACN 7/3, FA (0.5%)	LC/MS–MS	[72]
	Pure water	-	Sol-gel entrapment (TEOS)	-; 4.28 µg	Off-line SPE	1 L (eq 2.5 g)/ 0.5 g	-	ACN/water 7/3	ELISA, LC/MS	[73]
MC-LR, MC-RR, MC-YR	Real waters	-	Glutaraldehyde-silica; purified pAbs	-; 1.8 µg/g IS		20 mL (0.5% MeOH)/ 0.25 g	Water + water/MeOH 8/2	MeOH/water 8/2	ELISA, PP2A, LC/MS	[74]
MC-LR, MC-RR, MC-YR	Cyanobacteria, real waters	MeOH/water, 75/25; x0.75, PBS	pAbs	-		100 µL	PBS + water + MeOH/water 25/75	MeOH	LC/DAD, CE (MECK)	[75]
MC-LR, MC-RR, MC-YR, MC-LA	Algae and fish extracts, real waters	-; <15% MeOH	CNBr-Sepharose and silica; -	-		5–15 mL/ 0.1–0.2 g	PBS + water + MeOH/water 25/75	MeOH/water 8/2, AA (4%)	LC/UV	[76]
MC-LR, MC-RR, MC-YR, MC-LA	Real waters and blue green algae extracts	-	Sepharose CL-4B; pAbs (1 mg/mg sorbent)	-; 0.2 µg		10 mL/ 2 mg	PBS + water + water/MeOH 85/15	MeOH/water 80/20, AA (2%)	ELISA, LC/UV	[77]
MC-RR, MC-YR, MC-LR, MC-AR, MC-FR, MC-WR, MC-LA, MC-LF, MCYST-LW and other MC variants	Real waters	Concentrated, filtered	CNBr-Sepharose and silica; pAbs	-; 200 ng/IS (Sepharose); 135 ng/IS (silica)		-	Water/MeOH 75/25	MeOH/water (+AA) 8/2	LC/DAD, LC/MS	[78]
	Urine	-	Streptavidin-beads (2.5 µL); biotinylated Abs (0.5 µg)	-	dSPE	500 µL/ 2.5 µL	-	Water/ACN 7/3, FA (0.5%)	PP2A (inhibition assay)	[79]
Phycotoxins—Diarrheic shellfish poisoning (DSP) toxins
OA	Shellfish	MeOH, NaOH; dried extract in PBS	Protein G-magnetic beads; mAbs (1 mg/mg sorbent)	-	dSPE	1 mL/ 1 mg	PBS	MeOH	LC/MS–MS	[80]
OA	Algae extract	-; PBS/ACN 8/2	Silica; pAbs	-	Off-line SPE	-/125 mg	MeOH/water 3/7	MeOH-water 8/2	LC/Fluo, LC/MS	[81]
OA and derived form	Shellfish (hepatopancreas)	LLE; water/ACN 8/2	Glutaraldehyde-silica; pAbs	-; 16 µg/g IS		2 mL/ 125 mg	MeOH/water 7/3	PBS/ACN 7/3	LC/Fluo, LC/MS	[82]
OA, DTX-1 and DTX 2	Shellfish		-; Anti-AO mAbs	-		-	-	-	LC/Fluo	[83]
Mycotoxins
AFs (B₁, B₂, G₁, G₂), OTA, ZON, SMC, T-2	Feed samples	ACN/water/AA 80/18/2; x3, PBS	CNBr-Crystarose; 4 mAbs (5 mg each/g sorbent)	-; 0.13 µg/mg Abs (sum of toxin)	Off-line SPE	10 mL (eq 0.6 g)/ 0.3 ml	PBS	MeOH	LC/MS–MS	[84]
AFs (B₁, B₂, G₁, G₂), OTA, ZON, T-2	Peanuts, corn, wheat	ACN/water/AA 80/19/1; x3, PBS (≤ 20% ACN)	CNBr-Sepharose (1.3 g); mAbs (20 mg each)	-; 9 µg/mL IS (sum of toxin)		10 mL/ 0.1 mL	PBS		LC/MS–MS	[85]
AF B₁	Cereals, peanuts, vegetable oils, Chinese traditional food	MeOH/water 6/4; x6, water (<10% MeOH)	CNBr-Sepharose (1 g, 3.5 mL); mAbs (9.92 g)	99.8%; 260 ng/mL		30 mL (eq. 1 g)/ 1 mL	Water		LC/MS	[86]
α- and β-Amanitins	Urine	Filtration; x1.8, PB	CNBr-Sepharose 2 mL; pAbs (6.4 mg)			9 mL/ 2 mL	PB + water + acetone/water 95/5	Acetone/ MeOH 1/1	LC/MS	[87]
DON	Cereals	MeOH/water 8/2; x2, PBS (<10% MeOH)	CNBr-Sepharose (1 g, 3.5 mL); mAbs (30 mg)	95%; 9.67 nmol/mL		10 mL (eq. 0.5 g)/ 1 mL	Water + MeOH/water 1/9	MeOH	LC/MS–MS	[88]
DON, 3-ADON, 15-ADON, deepoxy-DON	Foods, feeds (aqueous extracts)	-	Abs entrapped in silica gel (TMOS); mAbs	-; 1 µg/mg immob. Abs		/1 g	1% MeOH	ACN/water 4/6	LC/UV	[89]
FUMs (B₁, B₂) B₃, OH-B₁)	Dried feed samples	-; 100 µL for 10 mg, buffer	Protein A/PS-DVB (POROS); serum/mL sorbent	-		100 µL/ 30 × 2.1 mm column	-	Water/MeOH 7/3, FA (2%)	LC/MS	[90]
FUMs (B₁, B₂, B₃)	Cereals	-	CNBr-Sepharose 4B (0.5 g); pAbs (1.27 mg/mL, 400 µL)	-		-	-	-	bioassay on cartridge	[91]
OTA	Beer	Degassed; x2, PBS, 1% PEG 6000	anti-IgG + CNBr-Sepharose (non-covalent bonding)	-		-	PBS, 0.05%Tween	OTA-HRP: competition	bioassay on cartridge	[92]
OTA	pure media	None	polyGMA-co-EGDMA monolith in a capillary; -	260 ng Ab/cm; 1.2 pmol OTA/cm	In-line SPE	10 µL/ 8.5 cm × 75 µm i.d	PBS+ borate buffer	MeOH	CE/LIF	[93]
T-2, HT-2	Maize, cherry	MeOH/water 6/4; x6, water, ≤ 10% MeOH	CNBr-Sepharose (1 g); mAbs (30 mg)	-; 3 µg/mL IS (for each toxin)	Off-line SPE	30 mL (eq 0.5 g)/1 mL, 10 × 0.8 mm column	-	MeOH	LC/MS–MS	[94]
ZER + 3 analogs	Bovine muscle	MeOH; x5, PBS	CNBr-Sepharose 2 g; mAbs (50 mg)	96.3%; 2.7 µg/mL gel		25 mL (eq 2.5 g)/1 mL, 10 × 0.8 mm column	PBS + water + water/MeOH 7/3	MeOH	GC/MS	[95]
ZON, DON, T-2, HT-2	Grain products	-	CNBr-Sepharose (0.2 g); DON Abs (1.25 mg), H-2/HT-2 Abs (0.2 mg), ZON Abs (0.3 mg)	100%; 198–281 ng (for each compound)		-	Water or PBS	MeOH	LC/MS–MS	[96]
ZON, DON, T-2, HT-2	Flour	-	Activated poly(GMA-co-DVB) µSpheres (0.3 g, 1 mL); DON Abs (1.25 mg), H-2/HT-2 Abs (0.2 mg), ZON Abs (0.3 mg)	-; 210–294 ng (for each compound)		/300 mg, 1 mL	-	-	LC/MS–MS	[97]
ZON, T-2, HT-2	Feed samples	ACN/H₂O, 8/2; x3, PBS	Anti-IgG-Sepharose (0.5 g, 1.8 mL); mAbs (ZON) and pAbs (T-2)	-		/0.2 g	-	-	bioassay, LC/MS–MS	[98]
Phycotoxins—Paralytic shellfish poisoning toxins
STX	Human urine	-	Protein G-magnetic beads (30 mg/mL); mAbs, (1 mg/mL)	15 µg/mg (theory); -	dSPE	100 µL/1.5 mg	PBS + water	ACN/water 1/1, FA (2.5%)	LC/MS–MS	[99]
STX, NEO	Shellfish	-	NH₂-coated hollow glass magnetic µSpheres; mAbs	5.8 mg/g; -		1 mL/25–100 mg	PBS	Glycine/HCl buffer	LC/Fluo	[100]
PSP toxins	Algae culture	PBS	NH₂-coated hollow glass magnetic µSpheres; mAbs	5.5 mg/g; -		1 mL/25–100 mg	PBS	Glycine/HCl buffer	LC/Fluo	[101]
Protein toxins
Abrin	Milk	-	Tosyl-activated magnetic beads (14.8 mg); mAbs against 4 epitopes (140 µg)	-	dSPE	500 µL/0.2 mg	PBS + Tween 0.05% + PBS + water	Trypsin digestion	LC/HRMS	[102]
Androctonus australis Hector	Venom		CNBr-Sepharose 2 g; purified pAbs (0.15 µmole)		Off-line SPE	/20 x1 cm column, 2 g	Tris HCl, NaCl	FA (pH 2.5), NaCl	UV	[103]
BoNT type A	Crude culture supernatant, food, environmental samples		Protein G-magnetic beads (3 µm), pAbs (BoNT A) and mAbs (ricin)		dSPE	500 µL/10–100 µL	HEPES	Trypsin digestion	LC/MS–MS	[104]
ETX	Milk, serum					500 µL/10–100 µL	HEPES	Trypsin digestion	LC/MS–MS	[105]
Ricin	Pure media (buffer + BSA)					500µL/100 µL	Ammonium acetate (pH 4)	RNA incubation	LC/MS on adenine	[106]
	Milk					500µL/5 µL	Buffer	5% FA or 0.1% TFA in water	Tryptic digestion + MALDI-MS or/and LC/MS	[107]
	Milk, apple juice, human serum, saliva					500µL/20 µL	PBS, Tween + water	ACN, TFA		[108]
	Serum		Streptavidin-magnetic beads; biotinylated mAbs	55 µg/mg; 16.5 µg/mg		500 µL/20 µL	PBS + water	TFA 0.1%		[109]
Ricin, SEB, BoTN A and B	Milk, orange and apple juices		M-280 tosyl- paramagnetic beads (250 µL); mAbs	-		200 µL/8 µL	PBS + water	TFA 0.1%		[110]
Ricin, SEB, ETX	Milk, human urine, plasma		M-280 tosyl- paramagnetic beads (250 µL); mAbs			1 mL/20 µL	PBS	Trypsin digestion	LC/HRMS (Q orbitrap)	[111]
Shigatoxin (protein) + analogs	Cell culture		CH-Sepharose 4B (2 g); purified pAbs (4 mg)		Off-line SPE	/2 g	Tris HCl + NaCl	Glycine (pH 2.7), NaCl 0.5 M	SDS Page	[112]
Staphilococcal enterotoxins A and E (proteins)	Dialyzed cell culture supernatant		Affigel 10 (agarose) 1 mL; mAbs (5 mg)		Off-line SPE	25 mL/1 mL	PB	AA, NaCl	UV	[113]

AA: acetic acid; APTES: aminopropyltriethoxysilane; BoTN: botulinium neurotoxin; CE: capillary electrophoresis; CNBr: cyanogen bromide; ETX: epsilon toxin; FA: formic acid, HRP: horseradish peroxidase; LLE, liquid–liquid extraction; NEO: neosaxitoxin; NHS: N-hydroxysuccinimide; PB: phosphate buffer (PBS: PB saline); PEG: polyethylene glycol; SEB: staphylococcal enterotoxin B; TEOS: tetraethoxysilane; TFA: trifluoroacetic acid; TTX: tetrodotoxin; ZER: zeranol. -: no data.